Methods and compounds for the treatment of immunologically-mediated skin disorders

ABSTRACT

Methods for the treatment of skin disorders, including psoriasis, atopic dermatitis, allergic contact dermatitis, alopecia areata, skin cancers, and related disorders, such as psoriatic arthritis are provided, such methods comprising administering a composition having antigenic and/or adjuvant properties. Compositions which may be usefully employed in the inventive methods include inactivated  M. vaccae  cells, delipidated and deglycolipidated  M. vaccae  cells,  M. vaccae  culture filtrate and compounds present in or derived therefrom, together with combinations of such compositions.

REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part of U.S. patentapplication Ser. No. 09/324,542, filed Jun. 2, 1999, which is acontinuation-in-part of U.S. patent application Ser. No. 08/997,080,filed Dec. 23, 1997, now U.S. Pat. No. 5,968,524.

TECHNICAL FIELD

[0002] This invention relates generally to the treatment by vaccinationor immunotherapy of skin disorders such as psoriasis, atopic dermatitis,allergic contact dermatitis, alopecia areata, the skin cancers basalcell carcinoma, squamous cell carcinoma and melanoma, and relateddisorders, such psoriatic arthritis. In particular, the invention isrelated to the use of compounds which are present in or have beenderived from Mycobacterium vaccae (M. vaccae) or from the culturefiltrate of M. vaccae.

BACKGROUND OF THE INVENTION

[0003] This invention deals with treatment of disorders of skin whichappear to be associated with factors that influence the balance ofthymus-derived (T) immune cells known as Th1 and Th2. These T cells areidentified by their cytokine secretion phenotype. A common feature oftreatment is the use of compounds prepared from M. vaccae which haveimmunomodulating properties that alter the balance of activities ofthese T cells as well as other immune cells.

[0004] Psoriasis is a common, chronic inflammatory skin disease whichcan be associated with various forms of arthritis in a minority ofpatients. The defect in psoriasis appears to be overly rapid growth ofkeratinocytes and shedding of scales from the skin surface. Drug therapyis directed at slowing down this process. The disease may becomemanifest at any age. Spontaneous remission is relatively rare, andlife-long treatment is usually necessary. Psoriasis produces chronic,scaling red patches on the skin surface. Psoriasis is a very visibledisease, it frequently affects the face, scalp, trunk and limbs. Thedisease is emotionally and physically debilitating for the patient,detracting significantly from the quality of life. Between one and threemillion individuals in the United States have psoriasis with nearly aquarter million new cases occurring each year. Conservative estimatesplace the costs of psoriasis care in the United States currently at $248million a year.

[0005] There are two major hypotheses concerning the pathogenesis ofpsoriasis. The first is that genetic factors determine abnormalproliferation of epidermal keratinocytes. The cells no longer respondnormally to external stimuli such as those involved in maintainingepidermal homeostasis. Abnormal expression of cell membrane cytokinereceptors or abnormal transmembrane signal transduction might underliecell hyperproliferation. Inflammation associated with psoriasis issecondary to the release of pro-inflammatory molecules fromhyperproliferative keratinocytes.

[0006] A second hypothesis is that T cells interacting withantigen-presenting cells in skin release pro-inflammatory andkeratinocyte-stimulating cytokines (Hancock, G. E. et al., J. Exp. Med.168:1395-1402, 1988). Only T cells of genetically predeterminedindividuals possess the capacity to be activated under suchcircumstances. The keratinocytes themselves may be theantigen-presenting cell. The cellular infiltrate in psoriatic lesionsshow an influx of CD4+ T cells and, more prominently, CD8+ T cells (Bos,J. D. et al., Arch. Dermatol. Res. 281:23-3, 1989; Baker, B. S., Br. J.Dermatol. 110:555-564, 1984).

[0007] As the majority (90%) of psoriasis patients have limited forms ofthe disease, topical treatments which include dithranol, tarpreparations, corticosteroids and the recently introduced vitamin D3analogues (calcipotriol, calcitriol) can be used. A minority (10%) ofpsoriasis patients have a more serious condition, for which a number ofsystemic therapeutic modalities are available. Specific systemictherapies include UVB, PUVA, methotrexate, vitamin A derivatives(acitretin) and immuno-suppressants such as Cyclosporin A. Theeffectiveness of Cyclosporin and FK-506 for treating psoriasis providessupport for the T cell hypothesis as the prime cause of the disease(Bos, J. D. et al., Lancet II: 1500-1502, 1989; Ackerman, C. et al., J.Invest. Dermatol. 96:536 [abstract], 1991).

[0008] About 5-10% of patients with psoriasis suffer from aninflammatory arthritis. While the pattern of arthritis is characteristicof psoriatic arthritis, active joint inflammation often does not occursimultaneously with the typical skin lesions of psoriasis; active jointdisease may be present while the skin disease is in relative remission.The inflammation in psoriatic arthritis (PsA) is also characterized byinfiltration of T cells into the synovium of affected joints.

[0009] Atopic dermatitis is a chronic pruritic inflammatory skin diseasewhich usually occurs in families with an hereditary predisposition forvarious allergic disorders such as allergic rhinitis and asthma. Atopicdermatitis occurs in approximately 10% of the general population. Themain symptoms are dry skin, dermatitis (eczema) localised mainly in theface, neck and on the flexor sides and folds of the extremitiesaccompanied by severe itching. It typically starts within the first twoyears of life. In about 90% of the patients this skin disease disappearsduring childhood but the symptoms can continue into adult life. It isone of the commonest forms of dermatitis world-wide. It is generallyaccepted that in atopy and in atopic dermatitis, a T cell abnormality isprimary and that the dysfunction of T cells which normally regulate theproduction of IgE is responsible for the excessive production of thisimmunoglobulin.

[0010] Allergic contact dermatitis is a common non-infectiousinflammatory disorder of the skin. In contact dermatitis, immunologicalreactions cannot develop until the body has become sensitised to aparticular antigen. Subsequent exposure of the skin to the antigen andthe recognition of these antigens by T cells result in the release ofvarious cytokines, proliferation and recruitment of T cells and finallyin dermatitis (eczema).

[0011] Only a small proportion of the T cells in a lesion of allergiccontact dermatitis are specific for the relevant antigen. Activated Tcells probably migrate to the sites of inflammation regardless ofantigen-specificity. Delayed-type hypersensitivity can only betransferred by T cells (CD4⁺ cells) sharing the MHC class II antigens.The ‘response’ to contact allergens can be transferred by T cellssharing either MHC class I (CD8⁺ cells) or class II (CD4⁺ cells)molecules (Sunday, M. E. et al., J. Immunol. 125:1601-1605, 1980).Keratinocytes can produce interleukin-1 which can facilitate the antigenpresentation to T cells. The expression of the surface antigenintercellular adhesion molecule-1 (ICAM-1) is induced both onkeratinocytes and endothelium by the cytokines tumor necrosis factor(TNF) and interferon-gamma (IFN-γ).

[0012] If the causes can be identified, removal alone will cure allergiccontact dermatitis. During active inflammation, topical corticosteroidsare useful. An inhibitory effect of cyclosporin has been observed indelayed-type hypersensitivity on the pro-inflammatory function(s) ofprimed T cells in vitro (Shidani, B. et al., Eur. J. Immunol.14:314-318, 1984). The inhibitory effect of cyclosporin on the earlyphase of T cell activation in mice has also been reported (Milon, G. etal., Ann. Immunol. (Inst. Pasteur) 135d:237-245, 1984).

[0013] Alopecia areata is a common hair disease, which accounts forabout 2% of the consultations at dermatological outpatient clinics inthe United States. The hallmark of this disease is the formation ofwell-circumscribed round or oval patches of nonscarring alopecia whichmay be located in any hairy area of the body. The disease may develop atany age. The onset is usually sudden and the clinical course is varied.

[0014] At present, it is not possible to attribute all or indeed anycase of alopecia areata to a single cause (Rook, A. and Dawber, R,Diseases of the Hair and Scalp, Blackwell Scientific Publications 1982:272-30). There are many factors that appear to be involved. Theseinclude genetic factors, atopy, association with disorders of supposedautoimmune etiology, Down's syndrome and emotional stress. Theprevalence of atopy in patients with alopecia areata is increased. Thereis evidence that alopecia areata is an autoimmune disease. This evidenceis based on consistent histopathological findings of a lymphocytic Tcell infiltrate in and around the hair follicles with increased numbersof Langerhans cells, the observation that alopecia areata will respondto treatment with immunomodulating agents, and that there is astatistically significant association between alopecia areata and a widevariety of autoimmune diseases (Mitchell, A. J. et al., J. Am. Acad.Dermatol. 11:763-775, 1984). Alopecia areata is associated with abnormalantibody production, which is believed to be associated with a Th2immune response.

[0015] Immunophenotyping studies on scalp biopsy specimens showsexpression of HLA-DR on epithelial cells in the presumptive cortex andhair follicles of active lesions of alopecia areata, as well as a T cellinfiltration with a high proportion of helper/inducer T cells in andaround the hair follicles, increased numbers of Langerhans cells and theexpression of ICAM-1 (Messenger, A. G. et al., J. Invest. Dermatol.85:569-576, 1985; Gupta, A. K. et al., J. Am. Acad. Dermatol.22:242-250, 1990).

[0016] The large variety of therapeutic modalities in alopecia areatacan be divided into four categories: (i) non-specific topical irritants;(ii) ‘immune modulators’ such as systemic corticosteroids and PUVA;(iii) ‘immune enhancers’ such as contact dermatitis inducers,cyclosporin and inosiplex; and (iv) drugs of unknown action such asminoxidil (Dawber, R. P. R. et al., Textbook of Dermatology, BlackwellScientific Publications, 5^(th) Ed, 1982:2533-2638). Non-specifictopical irritants such as dithranol may work through as yet unidentifiedmechanisms rather than local irritation in eliciting regrowth of hair.Topical corticosteroids may be effective but prolonged therapy is oftennecessary. Intralesional steroids have proved to be more effective buttheir use is limited to circumscribed patches of less active disease orto maintain regrowth of the eyebrows in alopecia totalis.Photochemotherapy has proved to be effective, possibly by changingfunctional subpopulations of T cells. Topical immunotherapy by means ofinduction and maintenance of allergic contact dermatitis on the scalpmay result in hair regrowth in as many as 70% of the patients withalopecia areata. Diphencyprone is a potent sensitiser free frommutagenic activity. Oral cyclosporin can be effective in the short term(Gupta, A. K. et al., J. Am. Acad. Dermatol. 22:242-250, 1990).Inosiplex, an immunostimulant, has been used with apparent effectivenessin an open trial. Topical 5% minoxidil solution has been reported to beable to induce some hair growth in patients with alopecia areata. Themechanism of action is unclear.

[0017] Carcinomas of the skin are a major public health problem becauseof their frequency and the disability and disfigurement that they cause.Carcinoma of the skin is principally seen in individuals in their primeof life, especially in fair skinned individuals exposed to large amountsof sunlight. The annual cost of treatment and time loss from workexceeds $250 million dollars a year in the United States alone. Thethree major types—basal cell cancer, squamous cell cancer, andmelanoma—are clearly related to sunlight exposure.

[0018] Basal cell carcinomas are epithelial tumours of the skin. Theyappear predominantly on exposed areas of the skin. In a recentAustralian study, the incidence of basal cell carcinomas was 652 newcases per year per 100,000 of the population. This compares with 160cases of squamous cell carcinoma or 19 of malignant melanoma (Giles, G.et al., Br. Med. J. 296:13-17, 1988). Basal cell carcinomas are the mostcommon of all cancers. Lesions are usually surgically excised. Alternatetreatments include retinoids, 5-fluorouracil, cryotherapy andradiotherapy. Alpha or gamma interferon have also been shown to beeffective in the treatment of basal cell carcinomas, providing avaluable alternative to patients unsuitable for surgery or seeking toavoid surgical scars (Cornell et al., J. Am. Acad. Dermatol. 23:694-700,1990; Edwards, L. et al., J. Am. Acad. Dermatol. 22:496-500, 1990).

[0019] Squamous cell carcinoma (SCC) is the second most common cutaneousmalignancy, and its frequency is increasing. There are an increasingnumber of advanced and metastatic cases related to a number ofunderlying factors. Currently, metastatic SCC contributes to over 2000deaths per year in the United States; the 5 year survival rate is 35%,with 90% of the metastases occurring by 3 years. Metastasis almostalways occurs at the first lymphatic drainage station. The need formedical therapy for advanced cases is clear. A successful medicaltherapy for primary SCC of the skin would obviate the need for surgicalexcision with its potential for scarring and other side effects. Thisdevelopment may be especially desirable for facial lesions.

[0020] Because of their antiproliferative and immunomodulating effectsin vitro, interferons (IFNs) have also been used in the treatment ofmelanoma (Kirkwood, J. M. et al., J. Invest. Dermatol. 95:180S-4S,1990). Response rates achieved with systemic IFN-α, in either high orlow dose, in metastatic melanoma were in the range 5-30%. Recently,encouraging results (30% response) were obtained with a combination ofIFN-α and DTIC. Preliminary observations indicate a beneficial effect ofIFN-α in an adjuvant setting in patients with high risk melanoma.Despite the low efficacy of IFN monotherapy in metastatic disease,several randomised prospective studies are now being performed with IFNsas an adjuvant or in combination with chemotherapy (McLeod, G. R. etal., J. Invest. Dermatol. 95: 185S-7S, 1990; Ho, V. C. et al., J.Invest. Dermatol. 22:159-76, 1990).

[0021] Of all the available therapies for treating cutaneous virallesions, only interferon possesses a specific antiviral mode of action,by reproducing the body's immune response to infection. Interferontreatment cannot eradicate the viruses however, although it may helpwith some manifestations of the infection. Interferon treatment is alsoassociated with systemic adverse effects, requires multiple injectionsinto each single wart and has a significant economic cost (Kraus, S. J.et al., Review of Infectious Diseases 2(6):S620-S632, 1990; Frazer, I.H., Current Opinion in Immunology 8(4):484-491, 1996).

[0022] Many compositions have been developed for topical application totreat skin disorders. Such topical treatments generally have limitedbeneficial effects. International Patent Publication WO 91/02542discloses treatment of chronic inflammatory disorders in which a patientdemonstrates an abnormally high release of IL-6 and/or TNF or in whichthe patient's IgG shows an abnormally high proportion of agalactosylIgG. Among the disorders mentioned in this publication are psoriasis,rheumatoid arthritis, mycobacterial disease, Crohn's disease, primarybiliary cirrhosis, sarcoidosis, ulcerative colitis, systemic lupuserythematosus, multiple sclerosis, Guillain-Barre syndrome, primarydiabetes mellitus, and some aspects of graft rejection. The therapeuticagent preferably comprises autoclaved M. vaccae administered byinjection in a single dose. This publication does not disclose anyclinical results.

[0023] Several other patents and publications disclose treatment ofvarious conditions by administering mycobacteria, including M. vaccae,or certain mycobacterial fractions. U.S. Pat. No. 4,716,038 disclosesdiagnosis of, vaccination against and treatment of autoimmune diseasesof various types, including arthritic diseases, by administeringmycobacteria, including M. vaccae. U.S. Pat. No. 4,724,144 discloses animmunotherapeutic agent comprising antigenic material derived from M.vaccae for treatment of mycobacterial diseases, especially tuberculosisand leprosy, and as an adjuvant to chemotherapy. International PatentPublication WO 91/01751 discloses the use of antigenic and/orimmunoregulatory material from M. vaccae as an immunoprophylactic todelay and/or prevent the onset of AIDS. International Patent PublicationWO 94/06466 discloses the use of antigenic and/or immunoregulatorymaterial derived from M. vaccae for therapy of HIV infection, with orwithout AIDS and with or without associated tuberculosis.

[0024] U.S. Pat. No. 5,599,545 discloses the use of mycobacteria,especially whole, inactivated M. vaccae, as an adjuvant foradministration with antigens which are not endogenous to M. vaccae. Thispublication theorises that the beneficial effect as an adjuvant may bedue to heat shock protein 65 (hsp 65). International Patent PublicationWO 92/08484 discloses the use of antigenic and/or immunoregulatorymaterial derived from M. vaccae for the treatment of uveitis.International Patent Publication WO 93/16727 discloses the use ofantigenic and/or immunoregulatory material derived from M. vaccae forthe treatment of mental diseases associated with an autoimmune reactioninitiated by an infection. International Patent Publication WO 95/26742discloses the use of antigenic and/or immunoregulatory material derivedfrom M. vaccae for delaying or preventing the growth or spread oftumors.

[0025]M. vaccae is apparently unique among known mycobacterial speciesin that heat-killed preparations retain vaccine and immunotherapeuticproperties. For example, M. bovis-BCG vaccines, used for vaccinationagainst tuberculosis, employ live strains. Heat-killed M. bovis BCG andM. tuberculosis have no protective properties when employed in vaccines.A number of compounds have been isolated from a range of mycobacterialspecies which have adjuvant properties. The effect of such adjuvants isessentially to stimulate a particular immune response mechanism againstan antigen from another species.

[0026] There are two general classes of compounds which have beenisolated from mycobacterial species that exhibit adjuvant properties.The first are water soluble wax D fractions (R. G. White, I. Bernstock,R. G. S. Johns and E. Lederer, Immunology, 1:54, 1958; U.S. Pat. No.4,036,953). The second are muramyl dipeptide-based substances (N-acetylglucosamine and N-glycolymuramic acid in approximately equimolaramounts) as described in U.S. Pat. Nos. 3,956,481 and 4,036,953. Thesecompounds differ from the delipidated and deglycolipidated M. vaccae(DD-M. vaccae) of the present invention in the following aspects oftheir composition:

[0027] 1. They are water-soluble agents, whereas DD-M. vaccae isinsoluble in aqueous solutions.

[0028] 2. They consist of a range of small oligomers of themycobacterial cell wall unit, either extracted from bacteria by varioussolvents, or digested from the cell wall by an enzyme. In contrast,DD-M. vaccae contains highly polymerised cell wall.

[0029] 3. All protein has been removed from their preparations bydigestion with proteolytic enzymes. The only constituents of theirpreparations are the components of the cell wall peptidoglycanstructure, namely alanine, glutamic acid, diaminopimelic acid, N-acetylglucosamine, and N-glycolylmuramic acid. In contrast, DD-M. vaccaecontains 50% w/w protein, comprising a number of distinct proteinspecies.

[0030] There thus remains a need in the art for effective compositionsand methods for the treatment of skin disorders that are inexpensive andcause few undesirable side effects.

SUMMARY OF INVENTION

[0031] Briefly stated, the present invention provides methods for thetreatment of the skin disorders, including psoriasis, atopic dermatitis,allergic contact dermatitis, alopecia areata, scleroderma and skincancers, such methods comprising administering an immunotherapeuticcomposition which is believed to have antigenic and/or adjuvantproperties. The immunotherapeutic compositions are preferablyadministered by intradermal injection.

[0032] In a first aspect, the inventive methods comprise administeringone or more doses of a composition including a component selected fromthe group consisting of inactivated M. vaccae cells, delipidated anddeglycolipidated M. vaccae cells, and components that are present in orderived from either M. vaccae cells or M. vaccae culture filtrate.Specific examples of components present in or derived from either M.vaccae cells or M. vaccae culture filtrate include polypeptides thatcomprise an immunogenic portion of an antigen, or a variant thereof,wherein the antigen includes a sequence selected from the groupconsisting of SEQ ID NO: 1-4, 9-16, 18-21, 23, 25, 26, 28, 29, 44, 45,47, 52-55, 63, 64, 70, 75, 89, 94, 98, 100-105, 109, 110, 112, 121, 124,125, 134, 135, 140, 141, 143, 145, 147, 152, 154, 156, 158, 160, 165,166, 170, 172, 174, 177, 178, 181, 182, 184, 186, 187, 192 and 194.

[0033] In a second aspect, the inventive methods comprise administeringa first dose of an immunotherapeutic composition at a first point intime and administering a second dose of the composition at a second,subsequent, point in time. Preferably, the multiple doses areadministered at intervals of about 2-4 weeks. In one embodiment,compositions which may be usefully employed in such methods comprise acomponent selected from the group consisting of inactivated M. vaccaecells, M. vaccae culture filtrate, delipidated and deglycolipidated M.vaccae cells, and constituents and combinations thereof In a secondembodiment, compositions for use in such methods comprise at least onecompound which is present in or derived from either M. vaccae cells orM. vaccae culture filtrate. Examples of such compounds includepolypeptides comprising an immunogenic portion of an antigen, or avariant thereof, wherein the antigen includes a sequence selected fromthe group consisting of SEQ ID NO: 1-4,9-16, 18-21, 23, 25, 26, 28, 29,44, 45, 47, 52-55, 63, 64, 70, 75, 89, 94, 98, 100-105, 109, 110, 112,121, 124, 125, 134, 135, 140, 141, 143, 145, 147, 152, 154, 156, 158,160, 165, 166, 170, 172, 174, 177, 178, 181, 182, 184, 186, 187, 192 and194.

[0034] Additional compositions which may be usefully employed in theinventive methods comprise a DNA molecule encoding one or more of theabove polypeptides. Compositions comprising a fusion protein, whereinthe fusion protein includes at least one of the above polypeptides,together with DNA molecules encoding such fusion proteins, may also beusefully employed in the methods of the present invention.

[0035] The compositions employed in the present invention mayadditionally include a non-specific immune response enhancer, oradjuvant. Such adjuvants may include M. vaccae culture filtrate,delipidated and deglycolipidated M. vaccae cells, or a polypeptidecomprising an immunogenic portion of an antigen, or a variant thereof,wherein said antigen includes a sequence provided in SEQ ID NO: 114, 117or 118.

[0036] The present invention further provides a method for treatingpsoriasis in a patient comprising administering a composition includinga component selected from the group consisting of inactivated M. vaccaecells; and delipidated and deglycolipidated M. vaccae cells, wherein thepatient has a PASI score of less than about 10 following treatment.

[0037] In yet further aspects, methods are provided for inhibiting a Th2immune response, and for treating skin disorders that are caused, atleast in part, by a Th2 immune response (for example, atopic dermatitis,allergic contact dermatitis, alopecia areata, skin disorders associatedwith systemic lupus erythematosus, and other antibody-mediated skindiseases) such methods comprising administering a composition comprisinginactivated M. vaccae cells, or delipidated and deglycolipidated M.vaccae cells. Methods are also provided for stimulating the productionof IL-10 and thereby inhibiting skin inflammation, such methodscomprising administering a composition comprising a component selectedfrom the group consisting of: inactivated M. vaccae cells, anddelipidated and deglycolipidated M. vaccae cells (DD-M. vaccae cells).

[0038] These and other aspects of the present invention will becomeapparent upon reference to the following detailed description andattached drawings. All references disclosed herein are herebyincorporated by reference in their entirety as if each was incorporatedindividually.

BRIEF DESCRIPTION OF THE DRAWINGS

[0039]FIG. 1 compares the stimulation of Interleukin 12 (IL-12)production in macrophages by different concentrations of heat-killed(autoclaved) M. vaccae, lyophilised M. vaccae, delipidated anddeglycolipidated M. vaccae, and M. vaccae glycolipids.

[0040]FIG. 2 compares the in vitro stimulation of interferon-gammaproduction in spleen cells from Severe Combined ImmunoDeficient (SCID)mice by different concentrations of heat-killed (autoclaved) M. vaccae,delipidated and deglycolipidated M. vaccae, and M. vaccae glycolipids.

[0041] FIGS. 3A(i)-(iv) illustrate the non-specific immune amplifyingeffects of 10 μg, 100 μg and 1 mg autoclaved M. vaccae and 75 μgunfractionated culture filtrates of M. vaccae, respectively.

[0042] FIGS. 3B(i) and (ii) illustrate the non-specific immuneamplifying effects of autoclaved M. vaccae, and delipidated anddeglycolipidated M. vaccae, respectively.

[0043]FIG. 3C(i) illustrates the non-specific immune amplifying effectsof whole autoclaved M. vaccae.

[0044] FIG. 3C(ii) illustrates the non-specific immune amplifyingeffects of soluble M. vaccae protein extracted with SDS from delipidatedand deglycolipidated M. vaccae.

[0045]FIG. 3C(iii) illustrates that the non-specific immune amplifyingeffects of the preparation of FIG. 3C(ii) are destroyed by treatmentwith the proteolytic enzyme Pronase.

[0046]FIG. 3D illustrates the non-specific immune amplifying effects ofheat-killed M. vaccae (FIG. 3D(i)), whereas a non-specific immuneamplifying effect was not seen with heat-killed preparations of M.tuberculosis (FIG. 3D(ii)), M. bovis BCG (FIG. 3D(iii)), M. phlei (FIG.3D(iv) or M. smegmatis (FIG. 3D(v)).

[0047] FIGS. 4A-E illustrate the effect of intranasal administration ofheat-killed M. vaccae, DD-M. vaccae or M. bovis BCG on the number ofeosinophils in BAL cells of mice sensitised and challenged withovalbumin. Control mice received PBS.

[0048]FIGS. 4A and B show the effect of administering either 10 or 1000μg of heat-killed M. vaccae (FIG. 4A), or 10, 100 or 200 μg of DD-M.vaccae (FIG. 4B) intranasally 4 weeks before intranasal challenge withovalbumin on eosinophil numbers in BAL cells.

[0049]FIGS. 4C and D show the effect of administering to mice either1000 μg of heat-killed M. vaccae (FIG. 4C) or 200 μg of DD-M. vaccae(FIG. 4D) intranasally one week before ovalbumin challenge. In

[0050]FIG. 4E, immunisation was with either 1 mg of heat-killed M.vaccae or 200 μg of DD-M. vaccae, given either intranasally (i.n.) orsubcutaneously (s.c.). In the same experiment, the effect ofimmunization with M. bovis BCG of the Pasteur (BCG-P) and Connought(BCG-C) strains prior to challenge was determined.

[0051]FIG. 5 shows the stimulation of IL-10 production in THP-1 cells byDD-M. vaccae.

DETAILED DESCRIPTION OF THE INVENTION

[0052] Effective vaccines that provide protection against infectiousmicroorganisms contain at least two functionally different components.The first is an antigen, which may be polypeptide or carbohydrate innature, and which is processed by macrophages and otherantigen-presenting cells and displayed for CD4⁺ T cells or for CD8⁺ Tcells. This antigen forms the “specific” target of an immune response.The second component of a vaccine is a non-specific immune responseamplifier, termed an adjuvant, with which the antigen is mixed or isincorporated into. An adjuvant amplifies either cell-mediated orantibody immune responses to a structurally unrelated compound orpolypeptide. Several known adjuvants are prepared from microbes such asBordetella pertussis, M. tuberculosis and M. bovis BCG. Adjuvants mayalso contain components designed to protect polypeptide antigens fromdegradation, such as aluminum hydroxide or mineral oil. While theantigenic component of a vaccine contains polypeptides that direct theimmune attack against a specific pathogen, such as M. tuberculosis, theadjuvant is often capable of broad use in many different vaccineformulations. Certain known proteins, such as bacterial enterotoxins,can function both as an antigen to elicit a specific immune response andas an adjuvant to enhance immune responses to unrelated proteins.

[0053] Certain pathogens, such as M. tuberculosis, as well as certaincancers, are effectively contained by an immune attack directed by CD4⁺and CD8⁺ T cells, known as cell-mediated immunity. Other pathogens, suchas poliovirus, also require antibodies, produced by B cells, forcontainment. These different classes of immune attack (T cell or B cell)are controlled by different subpopulations of CD4⁺ T cells, commonlyreferred to as Th1 and Th2 cells. A desirable property of an adjuvant isthe ability to selectively amplify the function of either Th1 or Th2populations of CD4⁺ T cells. Many skin disorders, including psoriasis,atopic dermatitis, alopecia, and skin cancers appear to be influenced bydifferences in the activity of these Th cell subsets.

[0054] Two types of Th cell subsets have been described in a murinemodel and are defined by the cytokines they release upon activation. TheTh1 subset secretes IL-2, IFN-γ and tumor necrosis factor, and mediatesmacrophage activation and delayed-type hypersensitivity response. TheTh2 subset releases IL-4, IL-5, IL-6 and IL-10, and stimulate B cellactivation. The Th1 and Th2 subsets are mutually inhibiting, so thatIL-4 inhibits Th1-type responses, and IFN-γ inhibits Th2-type responses.Similar Th1 and Th2 subsets have been found in humans, with release ofthe identical cytokines observed in the murine model. In particular, themajority of T-cell clones from atopic human lymphocytes resemble themurine Th2 cell that produces IL-4, whereas very few clones produceIFN-γ. Therefore, the selective expression of the Th2 subset withsubsequent production of IL-4 and decreased levels of IFN-γ-producingcells could lead to preferential enhancement of IgE production.

[0055] Inactivated M. vaccae and compounds derived from M. vaccae haveboth antigen and adjuvant properties which function to enhance Th1-typeimmune responses. The methods of the present invention employ one ormore of these antigen and adjuvant compounds from M. vaccae and/or itsculture filtrates to redirect immune activities of T cells in patients.Mixtures of such compounds are particularly effective in the methodsdisclosed herein. While it is well known that all mycobacteria containmany cross-reacting antigens, it is not known whether they containadjuvant compounds in common. As shown below, inactivated M. vaccae anda modified (delipidated and deglycolipidated) form of inactivated M.vaccae have been found to have adjuvant properties of the Th1-type whichare not shared by a number of other mycobacterial species. In addition,it has been found that inactivated M. vaccae and delipidated anddeglycolipidated M. vaccae (DD-M. vaccae) inhibit Th2 immune responses.DD-M. vaccae has also been shown to stimulate the production of IL-10and may therefore be effectively employed to inhibit skin inflammation.Furthermore, it has been found that M. vaccae produces compounds in itsown culture filtrate which amplify the immune response to M. vaccaeantigens also found in culture filtrate, as well as to antigens fromother sources.

[0056] The present invention provides methods for the immunotherapy ofskin disorders, including psoriasis, atopic dermatitis, alopecia, andskin cancers in patients, in which immunotherapeutic agents are employedto alter or redirect an existing state of immune activity by alteringthe function of T cells to a Th1-type of immune response, or to suppressa Th2 immune response. As used herein, a “patient” refers to anywarm-blooded animal, preferably a human. Compositions which may beusefully employed in the inventive methods comprise at least one of thefollowing components: inactivated M. vaccae cells; M. vaccae culturefiltrate; modified M. vaccae cells; and constituents and compoundspresent in or derived from M. vaccae and/or its culture filtrate. Asdetailed below, multiple administrations of such compositions,preferably by intradermal injection, have been shown to be highlyeffective in the treatment of psoriasis.

[0057] As used herein the term “inactivated M. vaccae” refers to M.vaccae that have either been killed by means of heat, as detailed belowin Examples 1 and 2, or subjected to radiation, such as ⁶⁰Cobalt at adose of 2.5 megarads. As used herein, the term “modified M. vaccae”includes delipidated M. vaccae cells, deglycolipidated M. vaccae cellsand M. vaccae cells that have been both delipidated anddeglycolipidated.

[0058] The preparation of delipidated and deglycolipidated-M. vaccae(DD-M. vaccae) and its chemical composition are described below inExample 1. As detailed below, the inventors have shown that removal ofthe glycolipid constituents from M. vaccae results in the removal ofmolecular components that stimulate interferon-gamma production innatural killer (NK) cells, thereby significantly reducing thenon-specific production of a cytokine that has numerous harmfulside-effects.

[0059] Compounds present in or derived from M. vaccae and/or from M.vaccae culture filtrate that may be usefully employed in the inventivemethods include polypeptides that comprise at least one immunogenicportion of an M. vaccae antigen, or a variant thereof, or at least oneadjuvant portion of an M. vaccae protein. In specific embodiments, suchpolypeptides comprise an immunogenic portion of an antigen, or a variantthereof, wherein the antigen includes a sequence selected from the groupconsisting of SEQ ID NO: 1-4,9-16, 18-21, 23, 25, 26, 28, 29, 44, 45,47, 52-55, 63, 64, 70, 75, 89, 94, 98, 100-105, 109, 110, 112, 121, 124,125, 134, 135, 140, 141, 143, 145, 147, 152, 154, 156, 158, 160, 165,166, 170, 172, 174, 177, 178, 181, 182, 184, 186, 187, 192 and 194.

[0060] As used herein, the term “polypeptide” encompasses amino acidchains of any length, including full length proteins (i.e. antigens),wherein the amino acid residues are linked by covalent peptide bonds.Thus, a polypeptide comprising an immunogenic portion of an antigen mayconsist entirely of the immunogenic portion, or may contain additionalsequences. The additional sequences may be derived from the native M.vaccae antigen or may be heterologous, and such sequences may (but neednot) be immunogenic. As detailed below, polypeptides of the presentinvention may be isolated from M. vaccae cells or culture filtrate, ormay be prepared by synthetic or recombinant means.

[0061] “Immunogenic”, as used herein, refers to the ability of apolypeptide to elicit an immune response in a patient, such as a human,or in a biological sample. In particular, immunogenic antigens arecapable of stimulating cell proliferation, interleukin-12 production orinterferon-γ production in biological samples comprising one or morecells selected from the group of T cells, NK cells, B cells andmacrophages, where the cells are derived from an individual previouslyexposed to tuberculosis. Exposure to an immunogenic antigen usuallyresults in the generation of immune memory such that upon re-exposure tothat antigen, an enhanced and more rapid response occurs.

[0062] Immunogenic portions of the antigens described herein may beprepared and identified using well known techniques, such as thosesummarised in Paul, Fundamental Immunology, 3d ed., Raven Press, 1993,pp. 243-247. Such techniques include screening polypeptide portions ofthe native antigen for immunogenic properties. The representativeproliferation and cytokine production assays described herein may beemployed in these screens. An immunogenic portion of a polypeptide is aportion that, within such representative assays, generates an immuneresponse (e.g., cell proliferation, interferon-γ production orinterleukin-12 production) that is substantially similar to thatgenerated by the full-length antigen. In other words, an immunogenicportion of an antigen may generate at least about 20%, preferably about65%, and most preferably about 100% of the proliferation induced by thefull-length antigen in the model proliferation assay described herein.An immunogenic portion may also, or alternatively, stimulate theproduction of at least about 20%, preferably about 65% and mostpreferably about 100%, of the interferon-y and/or interleukin-12 inducedby the full length antigen in the model assay described herein.

[0063] A M. vaccae adjuvant is a compound found in or derived from M.vaccae cells or M. vaccae culture filtrates which non-specificallystimulates immune responses. Adjuvants enhance the immune response toimmunogenic antigens and the process of memory formation. In the case ofM. vaccae antigens, these memory responses favor Th1-type immunity.Adjuvants are also capable of stimulating interleukin-12 production orinterferon-y production in biological samples comprising one or morecells selected from the group of T cells, NK cells, B cells andmacrophages, where the cells are derived from healthy individuals.Adjuvants may or may not stimulate cell proliferation. Such M. vaccaeadjuvants include, for example, the antigens of SEQ IDNO: 114, 117, 118.

[0064] The compositions which may be employed in the inventive methodsalso encompass variants of the described polypeptides. As used herein,the term “variant” covers any sequence which has at least about 40%,more preferably at least about 60%, more preferably yet at least about75% and most preferably at least about 90% identical residues (eithernucleotides or amino acids) to a sequence of the present invention. Thepercentage of identical residues is determined by aligning the twosequences to be compared, determining the number of identical residuesin the aligned portion, dividing that number by the total length of theinventive, or queried, sequence and multiplying the result by 100.

[0065] Polynucleotide or polypeptide sequences may be aligned, andpercentage of identical nucleotides in a specified region may bedetermined against another polynucleotide, using computer algorithmsthat are publicly available. Two exemplary algorithms for aligning andidentifying the similarity of polynucleotide sequences are the BLASTNand FASTA algorithms. The similarity of polypeptide sequences may beexamined using the BLASTP or FASTX algorithms. Both the BLASTN andBLASTP software are available on the NCBI anonymous FTP server(ftp://ncbi.nlm.nih.gov) under/blast/executables/. The BLASTN algorithmversion 2.0.4 [Feb. 24, 1998], set to the default parameters describedin the documentation and distributed with the algorithm, is preferredfor use in the determination of variants according to the presentinvention. The use of the BLAST family of algorithms, including BLASTNand BLASTP, is described at NCBI's website and in the publication ofAltschul, Stephen F., et al. (1997), “Gapped BLAST and PSI-BLAST: a newgeneration of protein database search programs”, Nucleic Acids Res.25:3389-3402. The computer algorithm FASTA is available on the Internet.Version 2.0u4, February 1996, set to the default parameters described inthe documentation and distributed with the algorithm, is preferred foruse in the determination of variants according to the present invention.The use of the FASTA algorithm is described in W. R. Pearson and D. J.Lipman, “Improved Tools for Biological Sequence Analysis,” Proc. Natl.Acad. Sci. USA 85:2444-2448 (1988) and W. R. Pearson, “Rapid andSensitive Sequence Comparison with FASTP and FASTA,” Methods inEnzymology 183:63-98 (1990). The use of the FASTX algorithm is describedin Pearson, W. R., Wood, T., Zhang, Z. and Miller, W., “Comparison ofDNA sequences with protein sequences,” Genomics 46(1):24-36 (1997).

[0066] The following running parameters are preferred for determinationof alignments and similarities using BLASTN that contribute to the Evalues and percentage identity: Unix running command: blastall -p blastn-d embldb -e 10-G 1-E 1 -r 2 -v 50 -b 50 -i queryseq -o results; andparameter default values:

[0067] -p Program Name [String]

[0068] -d Database [String]

[0069] -e Expectation value (E) [Real]

[0070] -G Cost to open a gap (zero invokes default behavior) [Integer]

[0071] -E Cost to extend a gap (zero invokes default behavior) [Integer]

[0072] -r Reward for a nucleotide match (blastn only) [Integer]

[0073] -v Number of one-line descriptions (V) [Integer]

[0074] -b Number of alignments to show (B) [Integer]

[0075] -i Query File [File In]

[0076] -o BLAST report Output File [File Out] Optional

[0077] For BLASTP the following running parameters are preferred:blastall -p blastp -d swissprotdb -e 10 -G 1 -E 1 -v 50 -b 50 -iqueryseq -o results

[0078] -p Program Name [String]

[0079] -d Database [String]

[0080] -e Expectation value (E) [Real]

[0081] -G Cost to open a gap (zero invokes default behavior) [Integer]

[0082] -E Cost to extend a gap (zero invokes default behavior) [Integer]

[0083] -v Number of one-line descriptions (v) [Integer]

[0084] -b Number of alignments to show (b) [Integer]

[0085] -I Query File [File In]

[0086] -o BLAST report Output File [File Out] Optional

[0087] The “hits” to one or more database sequences by a queriedsequence produced by BLASTN, BLASTP, FASTA, or a similar algorithm,align and identify similar portions of sequences. The hits are arrangedin order of the degree of similarity and the length of sequence overlap.Hits to a database sequence generally represent an overlap over only afraction of the sequence length of the queried sequence.

[0088] The BLASTN and FASTA algorithms also produce “Expect” values foralignments. The Expect value (E) indicates the number of hits one can“expect” to see over a certain number of contiguous sequences by chancewhen searching a database of a certain size. The Expect value is used asa significance threshold for determining whether the hit to a database,such as the preferred EMBL database, indicates true similarity. Forexample, an E value of 0.1 assigned to a hit is interpreted as meaningthat in a database of the size of the EMBL database, one might expect tosee 0.1 matches over the aligned portion of the sequence with a similarscore simply by chance. By this criterion, the aligned and matchedportions of the sequences then have a probability of 90% of being thesame. For sequences having an E value of 0.01 or less over aligned andmatched portions, the probability of finding a match by chance in theEMBL database is 1% or less using the BLASTN or FASTA algorithm.

[0089] According to one embodiment, “variant” polynucleotides, withreference to each of the polynucleotides of the present invention,preferably comprise sequences having the same number or fewer nucleicacids than each of the polynucleotides of the present invention andproducing an E value of 0.01 or less when compared to the polynucleotideof the present invention. That is, a variant polynucleotide is anysequence that has at least a 99% probability of being the same as thepolynucleotide of the present invention, measured as having an E valueof 0.01 or less using the BLASTN or FASTA algorithms set at the defaultparameters. According to a preferred embodiment, a variantpolynucleotide is a sequence having the same number or fewer nucleicacids than a polynucleotide of the present invention that has at least a99% probability of being the same as the polynucleotide of the presentinvention, measured as having an E value of 0.01 or less using theBLASTN or PASTA algorithms set at the default parameters.

[0090] Variant polynucleotide sequences will generally hybridize to therecited polynucleotide sequence under stringent conditions. As usedherein, “stringent conditions” refers to prewashing in a solution of 6×SSC, 0.2% SDS; hybridizing at 65° C., 6× SSC, 0.2% SDS overnight;followed by two washes of 30 minutes each in 1× SSC, 0.1% SDS at 65° C.and two washes of 30 minutes each in 0.2× SSC, 0.1% SDS at 65° C.

[0091] Polypeptide constituents and variants of the antigens andadjuvants present in or derived from M. vaccae or M. vaccae culturefiltrate may be isolated from M. vaccae or culture filtrate, or may begenerated by synthetic or recombinant means. Synthetic polypeptideshaving fewer than about 100 amino acids, and generally fewer than about50 amino acids, may be generated using techniques well known to those ofordinary skill in the art. For example, such polypeptides may besynthesized using any of the commercially available solid-phasetechniques, such as the Merrifield solid-phase synthesis method, whereamino acids are sequentially added to a growing amino acid chain. SeeMerrifield, J. Am. Chem. Soc. 85:2149-2146, 1963. Equipment forautomated synthesis of polypeptides is commercially available fromsuppliers such as Perkin Elmer/Applied BioSystems, Inc. (Foster City,Calif.), and may be operated according to the manufacturer'sinstructions. Variants of a native antigen or adjuvant may be preparedusing standard mutagenesis techniques, such as oligonucleotide-directedsite specific mutagenesis. Sections of the DNA sequence may also beremoved using standard techniques to permit preparation of truncatedpolypeptides, polypeptide fragments, and the like.

[0092] The polypeptides of the present invention may be altered ormodified, as is well known in the art, to confer desirable properties. Apolypeptide of the present invention may, for example, be conjugated toa signal (or leader) sequence at the N-terminal end of the protein whichco-translationally or post-translationally directs transfer of theprotein. The polypeptide may also be conjugated to a linker or othersequence for ease of synthesis, purification or identification of thepolypeptide (e.g., poly-His), or to enhance binding of the polypeptideto a solid support. For example, a polypeptide may be conjugated to animmunoglobulin Fc region. Other modifications may similarly be madewithout changing the activity of the polypeptide with respect totreatment of immunologically-mediated skin disorders. All such modifiedpolypeptides are within the scope of the present invention.

[0093] In general, M. vaccae antigens and adjuvants, and DNA sequencesencoding such antigens and adjuvants, may be prepared using any of avariety of procedures. For example, soluble antigens and adjuvants maybe isolated from M. vaccae culture filtrate as described below. Antigensor adjuvants may also be produced recombinantly by inserting a DNAsequence that encodes the antigen or adjuvant into an expression vectorand expressing the antigen or adjuvant in an appropriate host. Any of avariety of expression vectors known to those of ordinary skill in theart may be employed. Expression may be achieved in any appropriate hostcell that has been transformed or transfected with an expression vectorcontaining a DNA molecule that encodes recombinant polypeptide. Suitablehost cells include prokaryotes, yeast and higher eukaryotic cells.Preferably, the host cells employed are E. coli, yeast or a mammaliancell line such as COS or CHO. The DNA sequences expressed in this mannermay encode naturally occurring antigens, portions of naturally occurringantigens or adjuvants, or other variants thereof DNA sequences encodingM. vaccae antigens or adjuvants may be obtained by screening anappropriate M. vaccae cDNA or genomic DNA library for DNA sequences thathybridize to degenerate oligonucleotides derived from partial amino acidsequences of isolated soluble antigens or adjuvants. Suitable degenerateoligonucleotides may be designed and synthesized, and the screen may beperformed as described, for example, in Sambrook J, Fritsch E F andManiatis T, eds., Molecular Cloning: A Laboratory Manual, 2nd ed., ColdSpring Harbor Laboratory Press, Cold Spring Harbor: N.Y., 1989.. Asdescribed below, polymerase chain reaction (PCR) may be employed toisolate a nucleic acid probe from genomic DNA, or a cDNA or genomic DNAlibrary. The library screen may then be performed using the isolatedprobe.

[0094] DNA molecules encoding M. vaccae antigens may also be isolated byscreening an appropriate M. vaccae cDNA or genomic DNA expressionlibrary with anti-sera (e.g., rabbit or monkey) raised specificallyagainst M. vaccae antigens, as detailed below.

[0095] Regardless of the method of preparation, the antigens describedherein have the ability to induce an immunogenic response. Morespecifically, the antigens have the ability to induce cell proliferationand/or cytokine production (for example, interferon-γ and/orinterleukin-12 production) in T cells, NK cells, B cells or macrophagesderived from an M. tuberculosis-immune individual. A M.tuberculosis-immune individual is one who is considered to be resistantto the development of tuberculosis by virtue of having mounted aneffective T cell response to M. tuberculosis. Such individuals may beidentified based on a strongly positive (i.e., greater than about 10 mmdiameter induration) intradermal skin test response to tuberculosisproteins (PPD), and an absence of any symptoms of tuberculosisinfection. Among these immunogenic antigens, polypeptides havingsuperior therapeutic properties may be distinguished based on themagnitude of the responses in the assays described below.

[0096] Assays for cell proliferation or cytokine production in T cells,NK cells, B cell macrophages may be performed, for example, using theprocedures described below. The selection of cell type for use inevaluating an immune response to an antigen will depend on the desiredresponse. For example, interleukin-12 or interferon-γ production is mostreadily evaluated using preparations containing T cells, NK cells, Bcells and macrophages derived from individuals using methods well knownin the art. For example, a preparation of peripheral blood mononuclearcells (PBMCs) may be employed without further separation of componentcells. PBMCs may be prepared, for example, using density centrifugationthrough FiCol™ (Winthrop Laboratories, NY). T cells for use in theassays described herein may be purified directly from PBMCs.

[0097] In general, regardless of the method of preparation, thepolypeptides employed in the inventive methods are prepared insubstantially pure form. Preferably, the polypeptides are at least about80% pure, more preferably at least about 90% pure and most preferably atleast about 99% pure. In certain preferred embodiments, described indetail below, the substantially pure polypeptides are incorporated intopharmaceutical compositions or vaccines for use in one or more of themethods disclosed herein.

[0098] Fusion proteins comprising a first and a second inventivepolypeptide disclosed herein or, alternatively, a polypeptide disclosedherein and a known M. tuberculosis antigen, such as the 38 kDa antigendescribed in Andersen and Hansen, Infect. Immun. 57:2481-2488, 1989,together with variants of such fusion proteins, may also be employed inthe inventive methods. Such fusion proteins may include a linker peptidebetween the first and second polypeptides. A DNA sequence encoding sucha fusion protein is constructed using known recombinant DNA techniquesto assemble separate DNA sequences encoding the first and secondpolypeptides into an appropriate expression vector. The end of a DNAsequence encoding the first polypeptide is ligated, with or without apeptide linker, to the 5′ end of a DNA sequence encoding the secondpolypeptide so that the reading frames of the sequences are in phase topermit mRNA translation of the two DNA sequences into a single fusionprotein that retains the biological activity of both the first and thesecond polypeptides.

[0099] A peptide linker sequence may be employed to separate the firstand the second polypeptides by a distance sufficient to ensure that eachpolypeptide folds into its secondary and tertiary structures. Such apeptide linker sequence is incorporated into the fusion protein usingstandard techniques well known in the art. Suitable peptide linkersequences may be chosen based on the following factors: (1) theirability to adopt a flexible extended conformation; (2) their inabilityto adopt a secondary structure that could interact with functionalepitopes on the first and second polypeptides; and (3) the lack ofhydrophobic or charged residues that might react with the polypeptidefunctional epitopes. Preferred peptide linker sequences contain Gly, Asnand Ser residues. Other near neutral amino acids, such as Thr and Alamay also be used in the linker sequence. Amino acid sequences which maybe usefully employed as linkers include those disclosed in Maratea etal., Gene 40:39-46, 1985; Murphy et al., Proc. Natl. Acad. Sci. USA83:8258-8262, 1986; U.S. Pat. No. 4,935,233) and U.S. Pat. No.4,751,180. The linker sequence may be from 1 to about 50 amino acids inlength. Peptide linker sequences are not required when the first andsecond polypeptides have non-essential N-terminal amino acid regionsthat can be used to separate the functional domains and prevent stericinterference. The ligated DNA sequences encoding the fusion proteins arecloned into suitable expression systems using techniques known to thoseof ordinary skill in the art.

[0100] For use in the inventive methods, the inactivated M. vaccaecells; M. vaccae culture filtrate; modified M. vaccae cells; orcompounds present in or derived from M. vaccae and/or its culturefiltrate are generally present within a pharmaceutical composition or avaccine, with the pharmaceutical composition or vaccine being in a formsuitable for delivery via intradermal injection. Pharmaceuticalcompositions may comprise one or more components selected from the groupconsisting of inactivated M. vaccae cells, M. vaccae culture filtrate,modified M. vaccae cells, and compounds present in or derived from M.vaccae and/or its culture filtrate, together with a physiologicallyacceptable carrier. Vaccines may comprise one or more componentsselected from the group consisting of inactivated M. vaccae cells, M.vaccae culture filtrate, modified M. vaccae cells, and compounds presentin or derived from M. vaccae and/or its culture filtrate, together witha non-specific immune response amplifier. Such pharmaceuticalcompositions and vaccines may also contain other mycobacterial antigens,either, as discussed above, incorporated into a fusion protein orpresent within a separate polypeptide.

[0101] Alternatively, a vaccine or pharmaceutical composition for use inthe methods of the present invention may contain DNA encoding one ormore polypeptides as described above, such that the polypeptide isgenerated in situ. In such vaccines, the DNA may be present within anyof a variety of delivery systems known to those of ordinary skill in theart, including nucleic acid expression systems, bacterial and viralexpression systems. Appropriate nucleic acid expression systems containthe necessary DNA sequences for expression in the patient (such as asuitable promoter and terminator signal). Bacterial delivery systemsinvolve the administration of a bacterium (such as BacillusCalmette-Guerin) that expresses an immunogenic portion of thepolypeptide on its cell surface. In a preferred embodiment, the DNA maybe introduced using a viral expression system (e.g., vaccinia or otherpoxyirus, retrovirus, or adenovirus), which may involve the use of anon-pathogenic, or defective, replication competent virus. Techniquesfor incorporating DNA into such expression systems are well known in theart. The DNA may also be “naked,” as described, for example, in Ulmer etal., Science 259:1745-1749, 1993 and reviewed by Cohen, Science259:1691-1692.1993. The uptake of naked DNA may be increased by coatingthe DNA onto biodegradable beads, which are efficiently transported intothe cells.

[0102] While any suitable carrier known to those of ordinary skill inthe art may be employed in the pharmaceutical compositions of thisinvention, the type of carrier will vary depending on the mode ofadministration. For intradermal injection, the carrier preferablycomprises water, saline, alcohol, a fat, a lipid or a buffer.Biodegradable microspheres (e.g., polylactic galactide) may also beemployed as carriers for the pharmaceutical compositions and/or vaccinesof this invention. Suitable biodegradable microspheres are disclosed,for example, in U.S. Pat. Nos. 4,897,268 and 5,075,109. Any of a varietyof adjuvants may be employed in the vaccines of this invention tonon-specifically enhance the immune response.

[0103] While the frequency of administration, as well as dosage, willvary from individual to individual, multiple doses are preferablyadministered at intervals of about 2-4 weeks, more preferably atintervals of about 3 weeks and preferably by means of intradermalinjection. Alternate protocols may be appropriate for individualpatients. In some patients a booster dose may be administered on anannual basis.

[0104] The following examples are offered by way of illustration and arenot limiting.

EXAMPLE 1 Preparation and Immune Modulating Properties of Delipidatedand Deglycolipidated (DD-) M. vaccae

[0105] This example illustrates the processing of different constituentsof M. vaccae and their immune modulating properties.

[0106] Heat-Killed M. vaccae and M. vaccae Culture Filtrate

[0107]M. vaccae (ATCC Number 15483) was cultured in sterile Medium 90(yeast extract, 2.5 g/l; tryptone, 5 g/l; glucose 1 g/l) at 37° C. Thecells were harvested by centrifugation, and transferred into sterileMiddlebrook 7H9 medium (Difco Laboratories, Detroit, Mich., USA) withglucose at 37° C. for one day. The medium was then centrifuged to pelletthe bacteria, and the culture filtrate removed. The bacterial pellet wasresuspended in phosphate buffered saline at a concentration of 10 mg/ml,equivalent to 10¹⁰ M. vaccae organisms per ml. The cell suspension wasthen autoclaved for 15 min at 120° C. The culture filtrate was passagedthrough a 0.45 μM filter into sterile bottles.

[0108] Preparation of Delipidated and Deglycolipidated (DD-) M. vaccaeand Compositional Analysis

[0109] To prepare delipidated M. vaccae, the autoclaved M. vaccae waspelleted by centrifugation, the pellet washed with water, collectedagain by centrifugation and then freeze-dried. Freeze-dried M. vaccaewas treated with chloroform/methanol (2:1) for 60 mins at roomtemperature to extract lipids, and the extraction was repeated once. Thedelipidated residue from chloroform/methanol extraction was furthertreated with 50% ethanol to remove glycolipids by refluxing for twohours. The 50% ethanol extraction was repeated two times. The pooled 50%ethanol extracts were used as a source of M. vaccae glycolipids (seebelow). The residue from the 50% ethanol extraction was freeze-dried andweighed. The amount of delipidated and deglycolipidated M. vaccaeprepared was equivalent to 11.1% of the starting wet weight of M. vaccaeused. For bioassay, the delipidated and deglycolipidated M. vaccae,referred to as DD-M. vaccae, was resuspended in phosphate-bufferedsaline by sonication, and sterilized by autoclaving.

[0110] The compositional analyses of heat-killed M. vaccae and DD-M.vaccae are presented in Table 1. Major changes are seen in the fattyacid composition and amino acid composition of DD-M. vaccae as comparedto the insoluble fraction of heat-killed M. vaccae. The data presentedin Table 1 show that the insoluble fraction of heat-killed M. vaccaecontains 10% w/w of lipid, and the total amino acid content is 2750nmoles/mg, or approximately 33% w/w. DD-M. vaccae contains 1.3% w/w oflipid and 4250 nmoles/mg amino acids, which is approximately 51% w/w.TABLE 1 Compositional analyses of heat-killed M. vaccae and DD-M. vaccaeMONOSACCHARIDE COMPOSITION sugar alditol M. vaccae DD-M. vaccae Inositol3.2% 1.7% Ribitol* 1.7% 0.4% Arabinitol 22.7% 27.0% Mannitol 8.3% 3.3%Galactitol 11.5% 12.6% Glucitol 52.7% 55.2% FATTY ACID COMPOSITION Fattyacid M. vaccae DD-M. vaccae C14:0 3.9% 10.0% C16:0 21.1% 7.3% C16:114.0% 3.3% C18:0 4.0% 1.5% C18:1* 1.2% 2.7% C18:1w9 20.6% 3.1% C18:1w712.5% 5.9% C22:0 12.1% 43.0% C24:1* 6.5% 22.9%

[0111] The insoluble fraction of heat-killed M. vaccae contains 10% w/wof lipid, and DD-M. vaccae contains 1.3% w/w of lipid. AMINO ACIDCOMPOSITION nmoles/mg M. vaccae DD-M. vaccae ASP 231 361 THR 170 266 SER131 199 GLU 319 505 PRO 216 262 GLY 263 404 ALA 416 621 CYS* 24 26 VAL172 272 MET* 72 94 ILE 104 171 LEU 209 340 TYR 39 75 PHE 76 132 GlcNH2 56 HIS 44 77 LYS 108 167 ARG 147 272

[0112] The total amino acid content of the insoluble fraction ofheat-killed M. vaccae is 2750 nmoles/mg, or approximately 33% w/w. Thetotal amino acid content of DD-M. vaccae is 4250 nmoles/mg, orapproximately 51% w/w.

[0113]M. vaccae Glycolipids

[0114] The pooled 50% ethanol extracts described above were dried byrotary evaporation, redissolved in water and freeze-dried. The amount ofglycolipid recovered was 1.2% of the starting wet weight of M. vaccaeused. For bioassay, the glycolipids were dissolved in phosphate-bufferedsaline.

[0115] Stimulation of Cytokine Synthesis

[0116] Whole heat-killed M. vaccae and DD-M. vaccae were shown to havedifferent cytokine stimulation properties. The stimulation of a Th1immune response is enhanced by the production of interleukin-12 (IL-12)from macrophages. The ability of different M. vaccae preparations tostimulate IL-12 production was demonstrated as follows.

[0117] A group of C57BL/6J mice were injected intraperitoneally withDIFCO thioglycolate and, after three days, peritoneal macrophages werecollected and placed in cell culture with interferon-gamma for threehours. The culture medium was replaced and various concentrations ofwhole heat-killed M. vaccae, heat-killed M. vaccae which was lyophilisedand reconstituted for use in phosphate-buffered saline, DD-M. vaccae, orM. vaccae glycolipids were added. After three days at 37° C., theculture supernatants were assayed for the presence of IL-12 produced bymacrophages. As shown in FIG. 1, all the M. vaccae preparationsstimulated the production of IL-12 from macrophages.

[0118] By contrast, these same M. vaccae preparations were examined forthe ability to stimulate interferon-gamma production from Natural Killer(NK) cells. Spleen cells were prepared from Severe CombinedImmunodeficient (SCID) mice. These populations contain 75-80% NK cells.The spleen cells were incubated at 37° C. in culture with differentconcentrations of heat-killed M. vaccae, DD-M. vaccae, or M. vaccaeglycolipids. The data shown in FIG. 2 demonstrates that, whileheat-killed M. vaccae and M. vaccae glycolipids stimulate production ofinterferon-gamma, DD-M. vaccae stimulated relatively lessinterferon-gamma. The combined data from FIGS. 1 and 2 indicate that,compared with whole heat-killed M. vaccae, DD-M. vaccae is a betterstimulator of IL-12 than interferon gamma.

[0119] These findings demonstrate that removal of the lipid glycolipidconstituents from M. vaccae results in the removal of molecularcomponents that stimulate interferon-gamma from NK cells, therebyeffectively eliminating an important cell source of a cytokine that hasnumerous harmful side-effects. DD-M. vaccae thus retains Th1 immuneenhancing capacity by stimulating IL-12 production, but has lost thenon-specific effects that may come through the stimulation ofinterferon-gamma production from NK cells.

[0120] The adjuvant effect of DD-M. vaccae and a number of M. vaccaerecombinant antigens of the present invention was determined bymeasuring stimulation of IL-12 secretion from murine peritonealmacrophages. The cloning and purification of the recombinant proteinsare described in Examples 4 to 10. Recombinant proteins that exhibitedadjuvant properties are listed in Table 2. TABLE 2 Recombinant M. vaccaeproteins that exhibit adjuvant properties Mouse strain Antigen C57BL/6JBALB/cByJ GVs-3 + + GVc-4P + + GV-5 + + GV-5P + + GVc-7 + + GV-22B + NDGV-27 + + GV-27A + + GV-27B + + GV-42 + ND DD-M. vaccae + +

EXAMPLE 2 Effect of Intradermal Injection of Beat-Killed Mycobacteriumvaccae on Psoriasis in Human Patients

[0121] This example illustrates the effect of two intradermal injectionsof heat-killed Mycobacterium vaccae on psoriasis.

[0122]M. vaccae (ATCC Number 15483) was cultured in sterile Medium 90(yeast extract, 2.5 g/l; tryptone, 5 g/l; glucose, 1 g/l) at 37° C. Thecells were harvested by centrifugation, and transferred into sterileMiddlebrook 7H9 medium (Difco Laboratories, Detroit, Mich., USA) withglucose at 37° C. for one day. The medium was then centrifuged to pelletthe bacteria, and the culture filtrate removed. The bacterial pellet wasresuspended in phosphate buffered saline at a concentration of 10 mg/ml,equivalent to 10¹⁰ M. vaccae organisms per ml. The cell suspension wasthen autoclaved for 15 min at 120° C. and stored frozen at −20° C. Priorto use the M. vaccae suspension was thawed, diluted to a concentrationof 5 mg/ml in phosphate buffered saline, autoclaved for 15 min at 120°C. and 0.2 ml aliquoted under sterile conditions into vials for use inpatients.

[0123] Twenty four volunteer psoriatic patients, male and female, 15-61years old with no other systemic diseases were admitted to treatment.Pregnant patients were not included. The patients had PASI scores of12-35. The PASI score is a measure of the location, size and degree ofskin scaling in psoriatic lesions on the body. A PASI score of above 12reflects widespread disease lesions on the body. The study commencedwith a washout period of four weeks where the patients did not havesystemic anti-psoriasis treatment or effective topical therapy.

[0124] The 24 patients were then injected intradermally with 0.1 ml M.vaccae (equivalent to 500 μg). This was followed three weeks later witha second intradermal injection with the same dose of M. vaccae (500 μg).

[0125] Psoriasis was evaluated from four weeks before the firstinjection of heat-killed M. vaccae to twelve weeks after the firstinjection as follows:

[0126] A. The PASI scores were determined at −4, 0, 3, 6 and 12 weeks;

[0127] B. Patient questionnaires were completed at 0, 3, 6 and 12 weeks;and

[0128] C. Psoriatic lesions: each patient was photographed at 0, 3, 6, 9and 12 weeks.

[0129] The data shown in Table 3 describe the age, sex and clinicalbackground of each patient. TABLE 3 Patient Data in the Study of theEffect of M. vaccae in Psoriasis Code Duration of No. Patient Age/SexDisorder Admission PASI Score PS-001 D.C. 49/F 30 years 28.8 PS-002 E.S.41/F 4 months 19.2 PS-003 M.G. 24/F 8 months 18.5 PS-004 D.B. 54/M 2years 12.2 PS-005 C.E. 58/F 3 months 30.5 PS-006 M.G. 18/F 3 years 15.0PS-007 L.M. 27/M 3 years 19.0 PS-008 C.C 21/F 1 month 12.2 PS-009 E.G42/F 5 months 12.6 PS-010 J.G 28/M 7 years 19.4 PS-011 J.U 39/M 1 year15.5 PS-012 C.S 47/M 3 years 30.9 PS-013 H.B 44/M 10 years 30.4 PS-014N.J 41/M 17 years 26.7 PS-015 J.T 61/F 15 years 19.5 PS-016 L.P 44/M 5years 30.2 PS-017 E.N 45/M 5 years 19.5 PS-018 E.L 28/F 19 years 16.0PS-019 B.A 38/M 17 years 12.3 PS-020 P.P 58/F 1 year 13.6 PS-021 L.I27/F 8 months 22.0 PS-022 A.C 20/F 7 months 26.5 PS-023 C.A 61/F 10years 12.6 PS-024 F.T 39/M 15 years 29.5

[0130] All patients demonstrated a non-ulcerated, localised erythematoussoft indurated reaction at the injection site. No side effects werenoted, or complained of, by the patients. The data shown in Table 4,below, are the measured skin reactions at the injection site, 48 hours,72 hours and 7 days after the first and second injections of heat-killedM. vaccae. The data shown in Table 5, below, are the PASI scores of thepatients at the time of the first injection of M. vaccae (Day 0) and 3,6, 9, 12 and 24 weeks later.

[0131] It can clearly be seen that, by week 9 after the first injectionof M. vaccae, 16 of 24 patients showed a significant improvement in PASIscores. Seven of fourteen patients who have completed 24 weeks offollow-up remained stable with no clinical sign of redevelopment ofsevere disease. These results demonstrate the effectiveness of multipleintradermal injections of inactivated M. vaccae in the treatment ofpsoriasis. PASI scores below 10 reflect widespread healing of lesions.Histopathology of skin biopsies indicated that normal skin structure isbeing restored. Only one of the first seven patients who have completed28 weeks follow-up has had a relapse. TABLE 4 Skin Reaction Measurementsin Millimeter Time of Measurement First Injection Second Injection Code48 72 48 72 No. hours hours 7 days hours hours 7 days PS-001 12 × 10 12× 10 10 × 8  15 × 14 15 × 14 10 × 10 PS-002 18 × 14 20 × 18 18 × 14 16 ×12 18 × 12 15 × 10 PS-003 10 × 10 14 × 10 10 × 8  15 × 12 15 × 10 10 ×10 PS-004 14 × 12 22 × 18 20 × 15 20 × 20 20 × 18 14 × 10 PS-005 10 × 1013 × 10 DNR DNIR DNR DNR PS-006 10 × 8  10 × 10 6 × 4 12 × 10 15 × 15 10× 6  PS-007 15 × 15 18 × 16 12 × 10 15 × 13 15 × 12 12 × 10 PS-008 18 ×18 13 × 12 12 × 10 18 × 17 15 × 10 15 × 10 PS-009 13 × 13 18 × 15 12 ×8  15 × 13 12 × 12 12 × 7  PS-010 13 × 11 15 × 15 8 × 8 12 × 12 12 × 125 × 5 PS-011 17 × 13 14 × 12 12 × 11 12 × 10 12 × 10 12 × 10 PS-012 17 ×12 15 × 12 9 × 9 10 × 10 10 × 6  8 × 6 PS-013 18 × 11 15 × 11 15 × 10 15× 10 15 × 13 14 × 6  PS-014 15 × 12 15 × 11 15 × 10 13 × 12 14 × 10 8 ×5 PS-015 15 × 12 16 × 12 15 × 10 7 × 6 14 × 12 6 × 4 PS-016 6 × 5 6 × 66 × 5 8 × 8 9 × 8 9 × 6 PS-017 20 × 15 15 × 14 14 × 10 15 × 15 17 × 16DNR PS-018 14 × 10 10 × 8  10 × 8  12 × 12 10 × 10 10 × 10 PS-019 10 ×10 14 × 12 10 × 8  DNR 15 × 14 15 × 14 PS-020 15 × 12 15 × 15 12 × 15 15× 15 14 × 12 13 × 12 PS-021 15 × 12 15 × 12 7 × 4 11 × 10 11 × 10 11 ×8  PS-022 12 × 10 10 × 8  10 × 8  15 × 12 13 × 10 10 × 8  PS-023 13 × 1214 × 12 10 × 10 17 × 17 15 × 15 DNR PS-024 10 × 10 10 × 10 10 × 8  10 ×8  8 × 7 8 × 7

[0132] TABLE 5 Clinical Status of Patients after Injection of M. vaccae(PASI Scores) Code No. Day 0 Week 3 Week 6 Week 9 Week 12 Week 24 PS-00128.8 14.5 10.7 2.2 0.7 0 PS-002 19.2 14.6 13.6 10.9 6.2 0.6 PS-003 18.517.2 10.5 2.7 1.6 0 PS-004 12.2 13.4 12.7 7.0 1.8 0.2  PS-005* 30.5 DNR18.7 DNR DNR 0 PS-006 15.0 16.8 16.4 2.7 2.1 3.0 PS-007 19.0 15.7 11.65.6 2.2 0 PS-008 12.2 11.6 11.2 11.2 5.6 0 PS-009 12.6 13.4 13.9 14.415.3 13.0 PS-010 18.2 16.0 19.4 17.2 16.9 19.3 PS-011 17.2 16.9 16.716.5 16.5 15.5 PS-012 30.9 36.4 29.7 39.8** PS-013 19.5 19.2 18.9 17.814.7 17.8 PS-014 26.7 14.7 7.4 5.8 9.9 24.4*** PS-015 30.4 29.5 28.628.5 28.2 24.3 PS-016 30.2 16.8 5.7 3.2 0.8 PS-017 12.3 12.6 12.6 12.68.2 PS-018 16.0 13.6 13.4 13.4 13.2 PS-019 19.5 11.6 7.0 DNR DNR PS-02013.6 13.5 12.4 12.7 12.4 PS-021 22.0 20.2 11.8 11.4 15.5 PS-022 26.525.8 20.7 11.1 8.3 PS-023 12.6  9.2 6.6 5.0 4.8 PS-024 29.5 27.5 20.919.0 29.8

EXAMPLE 3 Effect of Intradermal Injection of Delipidated,Deglycolipidated Mycobacterium vaccae (DD-M. Vaccae) on Psoriasis inPatients

[0133] This example illustrates the effect of two intradermal injectionsof DD-M. vaccae on psoriasis.

[0134] Seventeen volunteer psoriatic patients, male and female, 18-48years old with no other systemic diseases were admitted to treatment.Pregnant patients were not included. The patients had PASI scores of12-30. As discussed above, the PASI score is a measure of the location,size and degree of skin scaling in psoriatic lesions on the body. A PASIscore of above 12 reflects widespread disease lesions on the body. Thestudy commenced with a washout period of four weeks where the patientsdid not have systemic anti-psoriasis treatment or effective topicaltherapy. The 17 patients were then injected intradermally with 0.1 mlDD-M. vaccae (equivalent to 100 μg). This was followed three weeks laterwith a second intradermal injection with the same dose of DD-M. vaccae(100 μg).

[0135] Psoriasis was evaluated from four weeks before the firstinjection of M. vaccae to 48 weeks after the first injection as follows:

[0136] A. the PASI scores were determined at −4, 0, 3, 6, 12, 24, 36 and48 weeks;

[0137] B. patient questionnaires were completed at 0, 3, 6, 9 and 12weeks and thereafter every 4 weeks; and

[0138] C. psoriatic lesions: each patient was photographed at 0, 3 weeksand thereafter at various intervals.

[0139] The data shown in Table 6 describe the age, sex and clinicalbackground of each patient. TABLE 6 Patient Data in the Study of theEffect of DD-M. vaccae in Psoriasis Code Duration of No. Patient Age/SexDisorder Admission PASI Score PS-025 A.S 25/F 2 years 12.2 PS-026 M.B45/F 3 months 14.4 PS-027 A.G 34/M 14 years 24.8 PS-028 E.M 31/M 4 years18.2 PS-029 A.L 44/M 5 months 18.6 PS-030 V.B 42/M 5 years 21.3 PS-031R.A 18/M 3 months 13.0 PS-032 42/M 23 years 30.0 PS-033 37/F 27 years15.0 PS-034 42/M 15 years 30.4 PS-035 35/M 6 years 13.2 PS-036 43/M 6years 19.5 PS-037 35/F 4 years 12.8 PS-038 44/F 7 months 12.6 PS-03920/F 1 year 16.1 PS-040 28/F 8 months 25.2 PS-041 48/F 10 years 20.0

[0140] All patients demonstrated a non-ulcerated, localised erythematoussoft indurated reaction at the injection site. No side effects werenoted, or complained of by the patients. The data shown in Table 7 arethe measured skin reactions at the injection site, 48 hours, 72 hoursand 7 10 days after the first injection of DD-M. vaccae, and 48 hoursand 72 hours after the second injection. TABLE 7 Skin ReactionMeasurements in Millimeters Time of Measurement First Injection SecondInjection Code No. 48 hours 72 hours 7 days 48 hours 72 hours PS-025 8 ×8 8 × 8 3 × 2 10 × 10 10 × 10 PS-026 12 × 12 12 × 12 8 × 8 DNR 14 × 14PS-027 9 × 8 10 × 10 10 × 8  9 × 5 9 × 8 PS-028 10 × 10 10 × 10 10 × 8 10 × 10 10 × 10 PS-029 8 × 6 8 × 6 5 × 5 8 × 8 8 × 8 PS-030 14 × 12 14 ×14 10 × 10 12 × 10 12 × 10 PS-031 10 × 10 12 × 12 10 × 6  14 × 12 12 ×10

[0141] The data shown in Table 8 are the PASI scores of the 17 patientsat the time of the first injection of DD-M. vaccae (Day 0), then 3,6,12,24, 36 and 48 weeks later, when available. TABLE 8 Clinical Status ofPatients after Injection of DD-M. vaccae (PASI Scores) Code Repeat No.Day 0 Week 3 Week 6 Week 12 Week 24 Week 36 Week 48 treatment PS-02512.2 4.1 1.8 1.4 1.7 0.2 15.8 Wk 48 PS-026 14.4 11.8 6.0 6.9 1.4 0.4PS-027 24.8 23.3 18.3 9.1 10.6 7.5 1.9 PS-028 18.2 24.1 28.6* PS-02918.6 9.9 7.4 3.6 0.8 0 0 PS-030 21.3 15.7 13.9 16.5 18.6 5.8 1.7 PS-03113.0 5.1 2.1 1.6 0.3 0 0 PS-032 30.0 28.0 20 12.4 20.4 19.0 21.5 Wk 44PS-033 19.0 12.6 5.9 4.0 12.6 21.1(wk 40) 7.1(wk 52) Wk 20 PS-034 30.431.2 31.6 32.4 25.5 33.0 Wk 20 PS-035 13.2 11.6 10.6 1.6 1.4(wk 20) 1.0PS-036 19.5 18.0 18.0 16.8 18.0 10.2 Wk 20, 32 PS-037 12.8 13.1 1.2 0 00 PS-038 12.6 12.6 12.7 10.0 Wk 12 PS-039 16.1 17.9 18.3 17.0 Wk 12PS-040 25.2 3.9 0.5 PS-041 20.0 12.7 0.8

[0142] These results show the significant improvement in PASI scores in16 patients after injection with DD-M. vaccae. One patient dropped outof the study at 12 weeks with the diagnosis of exfoliativedermatitis/psoriasis. Patients that relapsed received a second or thirdinjection of DD-M. vaccae at the time indicated in Table 8.

[0143] At 6 weeks follow-up (n=17), the PASI score improved by >50% in 9of 17 (53%) patients. At 12 weeks follow up (n=14), the PASI scoreimproved by >50% in 9 of 14 (64.3%) patients. Seven of these patientsshowed significant clinical improvement with reduction in PASI score toless than 8. At 24 weeks follow up (n=12), the PASI score improvedby >50% in 7 of 12 (58%) patients and at 48 weeks follow up (n=7), thePASI score improved by >50% in 5 of 7 (71%) patients. Again, four ofthese patients showed significant clinical improvement with reduction inPASI score to less than 2.

[0144] Local injection of DD-M. vaccae resulted in clearing of skinlesions at distant sites, thus indicating a systemic effect, andsuggesting that the systemic regulatory effects of treatment with DD-M.vaccae may be effective in reducing inflammation in the joints ofpatients with psoriatic arthritis.

EXAMPLE 4 The Non-Specific Immune Amplifying Properties of Heat-KilledM. vaccae, M. vaccae Culture Filtrate and DD-M. vaccae

[0145] This example illustrates the non-specific immune amplifying or‘adjuvant’ properties of whole heat-killed M. vaccae, DD-M. vaccae andM. vaccae culture filtrate.

[0146]M. vaccae bacteria was cultured, pelleted and autoclaved asdescribed in Example 1. Culture filtrates of live M. vaccae refer to thesupernatant from 24 h cultures of M. vaccae in 7H9 medium with glucose.DD-M. vaccae was prepared as described in Example 2.

[0147] Killed M. vaccae, DD-M. vaccae and M. vaccae culture filtratewere tested for adjuvant activity in the generation of cytotoxic T cellimmune response to ovalbumin, a structurally unrelated protein, in themouse. This anti-ovalbumin-specific cytotoxic response was detected asfollows. Groups of C57BL/6J mice were immunised by the intraperitonealinjection of 100 μg of ovalbumin with the following test adjuvants:heat-killed M. vaccae; DD-M. vaccae; DD-M. vaccae with proteinsextracted with SDS; the SDS protein extract treated with Pronase (anenzyme which degrades protein); and either heat-killed M. vaccae,heat-killed M. bovis BCG, M. phlei, M. smegmatis or M. vaccae culturefiltrate. After 10 days, spleen cells were stimulated in vitro for afurther 6 days with E.G7 cells which are EL4 cells (a C57BL/6J-derived Tcell lymphoma) transfected with the ovalbumin gene and thus expressovalbumin. The spleen cells were then assayed for their ability to killnon-specifically EL4 target cells or to kill specifically the E.G7ovalbumin expressing cells. Killing activity was detected by the releaseof Chromium with which the EL4 and E.G7 cells have been labelled (100mCi per 2×10⁶), prior to the killing assay. Killing or cytolyticactivity is expressed as % specific lysis using the formula:$\frac{{{cpm}\quad {in}\quad {test}\quad {cultures}} - {{cpm}\quad {in}\quad {control}\quad {cultures}}}{{{total}\quad {cpm}}\quad - {{cpm}\quad {in}\quad {control}\quad {cultures}}} \times 100\quad \%$

[0148] It is generally known that ovalbumin-specific cytotoxic cells aregenerated only in mice immunised with ovalbumin with an adjuvant but notin mice immunised with ovalbumin alone.

[0149] The diagrams that make up FIG. 3 show the effect of various M.vaccae derived adjuvant preparations on the generation of cytotoxic Tcells to ovalbumin in C57BL/6J mice. As shown in FIG. 3A, cytotoxiccells were generated in mice immunised with (i) 10 μg, (ii) 100 μg or(iii) 1 mg of autoclaved M. vaccae or (iv) 75 μg of M. vaccae culturefiltrate. FIG. 3B shows that cytotoxic cells were generated in miceimmunised with (i) 1 mg whole autoclaved M. vaccae or (ii) 100 μg DD-M.vaccae. As shown in FIG. 3C(i), cytotoxic cells were generated in miceimmunised with 1 mg heat-killed M. vaccae; FIG. 3C(ii) shows the activematerial in M. vaccae soluble proteins extracted with SDS from DD-M.vaccae. FIG. 3C(iii) shows that active material in the adjuvantpreparation of FIG. 3C(ii) was destroyed by treatment with theproteolytic enzyme Pronase. By way of comparison, 100 μg of theSDS-extracted proteins had significantly stronger immune-enhancingability (FIG. 3C(ii)) than did 1 mg heat-killed M. vaccae (FIG. 3C(i)).

[0150] Mice immunised with 1 mg heat-killed M. vaccae (FIG. 3D(i))generated cytotoxic cells to ovalbumin, but mice immunised separatelywith 1 mg heat-killed M. tuberculosis (FIG. 3D(ii)), 1 mg M. bovis BCG(FIG. 3D(iii)), 1 mg M. phlei (FIG. 3D(iv)), or 1 mg M. smegmatis (FIG.3D(v)) failed to generate cytotoxic cells.

[0151] The significance of these findings is that heat-killed M. vaccaeand DD-M. vaccae have adjuvant properties not seen in othermycobacteria. Further, delipidation and deglycolipidation of M. vaccaeremoves an NK cell-stimulating activity but does not result in a loss ofT cell-stimulating activity.

[0152] In subsequent studies, more of the SDS-extracted proteinsdescribed above were prepared by preparative SDS-PAGE on a BioRad PrepCell (Hercules, Calif.). Fractions corresponding to molecular weightranges were precipitated by trichloroacetic acid to remove SDS beforeassaying for adjuvant activity in the anti-ovalbumin-specific cytotoxicresponse assay in C57BL/6J mice as described above. The adjuvantactivity was highest in the 60-70 kDa fraction. The most abundantprotein in this size range was purified by SDS-PAGE blotted on to apolyinylidene difluoride (PVDF) membrane and then sequenced. Thesequence of the first ten amino acid residues is provided in SEQ IDNO:76. Comparison of this sequence with those in the gene bank asdescribed above, revealed homology to the heat shock protein 65 (GroEL)gene from M. tuberculosis, indicating that this protein is an M. vaccaemember of the GroEL family.

[0153] An expression library of M. vaccae genomic DNA in BamHI-lambdaZAP-Express (Stratagene) was screened using sera from cynomolgousmonkeys immunised with M. tuberculosis secreted proteins prepared asdescribed above. Positive plaques were identified using a colorimetricsystem. These plaques were re-screened until plaques were pure followingstandard procedures. pBK-CMV phagemid 2-1 containing an insert wasexcised from the lambda ZAP-Express (Stratagene) vector in the presenceof ExAssist helper phage following the manufacturer's protocol. The basesequence of the 5′ end of the insert of this clone, hereinafter referredto as GV-27, was determined using Sanger sequencing with fluorescentprimers on Perkin Elmer/Applied Biosystems Division automatic sequencer.The determined nucleotide sequence of the partial M. vaccaeGroEL-homologue clone GV-27 is provided in SEQ ID NO:77 and thepredicted amino acid sequence in SEQ ID NO:78. This clone was found tohave homology to M. tuberculosis GroEL.

[0154] A partial sequence of the 65 kDa heat shock protein of M. vaccaehas been published by Kapur et al. (Arch. Pathol. Lab. Med. 119:131-138,1995). However, this sequence did not overlap with the GV-27 sequenceprovided herein. The nucleotide sequence of the Kapur et al. fragment isshown in SEQ ID NO:79 and the predicted amino acid sequence in SEQ IDNO:80.

[0155] In subsequent studies, an extended DNA sequence (full-lengthexcept for the predicted 51 terminal residues) for GV-27 was obtained(SEQ ID NO: 113). The corresponding predicted amino acid sequence isprovided in SEQ ID NO: 114. Further studies led to the isolation of thefull-length DNA sequence for GV-27 (SEQ ID NO: 159). The correspondingpredicted amino acid sequence is provided in SEQ ID NO: 160. Thissequence shows 93.7% identity to the M. tuberculosis GroEL sequence. Twopeptide fragments, comprising the N-terminal sequence (hereinafterreferred to as GV-27A) and the carboxy terminal sequence of GV-27(hereinafter referred to as GV-27B) were prepared using techniques wellknown in the art. The nucleotide sequences for GV-27A and GV-27B areprovided in SEQ ID NO: 115 and 116, respectively, with the correspondingamino acid sequences being provided in SEQ ID NO: 117 and 118.Subsequent studies led to the isolation of an extended DNA sequence forGV-27B. This sequence is provided in SEQ ID NO: 161, with thecorresponding amino acid sequence being provided in SEQ ID NO: 162. Thesequence of GV-27A shows 95.8% identity to the published M. tuberculosisGroEL sequence and contains the M. vaccae sequence of Kapur et al.discussed above. The sequence of GV-27B is about 92.2% identical to thepublished M. tuberculosis sequence.

[0156] Following the same protocol as for the isolation of GV-27,pBK-CMV phagemid 3-1 was isolated. The antigen encoded by this DNA wasnamed GV-29. The determined nucleotide sequences of the 5′ and 3′ endsof the gene are provided in SEQ ID NOS: 163 and 164, respectively, withthe predicted corresponding amino acid sequences being provided in SEQID NOS: 165 and 166 respectively. GV-29 showed homology to yeast ureaamidolyase. The DNA encoding GV-29 was sub-cloned into the vector pET16(Novagen, Madison, Wis.) for expression and purification according tostandard protocols.

EXAMPLE 5 Purification and Characterization of Polypeptides from M.vaccae Culture Filtrate

[0157] This example illustrates the preparation of M. vaccae solubleproteins from culture filtrate. Unless otherwise noted, all percentagesin the following example are weight per volume.

[0158]M. vaccae (ATCC Number 15483) was cultured in sterile Medium 90 at37° C. The cells were harvested by centrifugation, and transferred intosterile Middlebrook 7H9 medium with glucose at 37° C. for one day. Themedium was then centrifuged (leaving the bulk of the cells) and filteredthrough a 0.45 μm filter into sterile bottles.

[0159] The culture filtrate was concentrated by lyophilization, andredissolved in MilliQ water. A small amount of insoluble material wasremoved by filtration through a 0.45 m membrane. The culture filtratewas desalted by membrane filtration in a 400 ml Amicon stirred cellwhich contained a 3,000 Da molecular weight cut-off (MWCO) membrane. Thepressure was maintained at 50 psi using nitrogen gas. The culturefiltrate was repeatedly concentrated by membrane filtration and dilutedwith water until the conductivity of the sample was less than 1.0 mS.This procedure reduced the 20 l volume to approximately 50 ml. Proteinconcentrations were determined by the Bradford protein assay (Bio-Rad,Hercules, Calif., USA).

[0160] The desalted culture filtrate was fractionated by ion exchangechromatography on a column of Q-Sepharose (Pharmacia Biotech, Uppsala,Sweden) (16×100 mm) equilibrated with 10 mM Tris HCl buffer pH 8.0.Polypeptides were eluted with a linear gradient of NaCl from 0 to 1.0 Min the above buffer system. The column eluent was monitored at awavelength of 280 nm.

[0161] The pool of polypeptides eluting from the ion exchange column wasconcentrated in a 400 ml Amicon stirred cell which contained a 3,000 DaMWCO membrane. The pressure was maintained at 50 psi using nitrogen gas.The polypeptides were repeatedly concentrated by membrane filtration anddiluted with 1% glycine until the conductivity of the sample was lessthan 0.1 mS.

[0162] The purified polypeptides were then fractionated by preparativeisoelectric focusing in a Rotofor device (Bio-Rad, Hercules, Calif.,USA). The pH gradient was established with a mixture of Ampholytes(Pharmacia Biotech) comprising 1.6% pH 3.5-5.0 Ampholytes and 0.4% pH5.0-7.0 Ampholytes. Acetic acid (0.5 M) was used as the anolyte, and 0.5M ethanolamine as the catholyte. Isoelectric focusing was carried out at12W constant power for 6 hours, following the manufacturer'sinstructions. Twenty fractions were obtained.

[0163] Fractions from isoelectric focusing were combined, and thepolypeptides were purified on a Vydac C4 column (Separations Group,Hesperia, Calif., USA) 300 Angstrom pore size, 5 micron particle size(10×250 mm). The polypeptides were eluted from the column with a lineargradient of acetonitrile (0-80% v/v) in 0.05% (v/v) trifluoroacetic acid(TFA). The flow-rate was 2.0 ml/min and the HPLC eluent was monitored at220 nm. Fractions containing polypeptides were collected to maximize thepurity of the individual samples.

[0164] Relatively abundant polypeptide fractions were rechromatographedon a Vydac C4 column (Separations Group) 300 Angstrom pore size, 5micron particle size (4.6×250 mm). The polypeptides were eluted from thecolumn with a linear gradient from 20-60% (v/v) of acetonitrile in 0.05%(v/v) TFA at a flow-rate of 1.0 ml/min. The column eluent was monitoredat 220 nm. Fractions containing the eluted polypeptides were collectedto maximise the purity of the individual samples. Approximately 20polypeptide samples were obtained and they were analysed for purity on apolyacrylamide gel according to the procedure of Laemmli (Laemmli, U.K., Nature 277:680-685, 1970).

[0165] The polypeptide fractions which were shown to contain significantcontamination were further purified using a Mono Q column (PharmaciaBiotech) 10 micron particle size (5×50 mm) or a Vydac Diphenyl column(Separations Group) 300 Angstrom pore size, 5 micron particle size(4.6×250 mm). From a Mono Q column, polypeptides were eluted with alinear gradient from 0-0.5 M NaCl in 10 mM Tris.HCl pH 8.0. From a VydacDiphenyl column, polypeptides were eluted with a linear gradient ofacetonitrile (20-60% v/v) in 0.1% TFA. The flow-rate was 1.0 ml/min andthe column eluent was monitored at 220 nm for both columns. Thepolypeptide peak fractions were collected and analysed for purity on a15% polyacrylamide gel as described above.

[0166] For sequencing, the polypeptides were individually dried ontoBiobrene™ (Perkin Elmer/Applied BioSystems Division, Foster City,Calif.)-treated glass fiber filters. The filters with polypeptide wereloaded onto a Perkin Elmer/Applied BioSystems Procise 492 proteinsequencer and the polypeptides were sequenced from the amino terminalend using traditional Edman chemistry. The amino acid sequence wasdetermined for each polypeptide by comparing the retention time of thePTH amino acid derivative to the appropriate PTH derivative standards.

[0167] Internal sequences were also determined on some antigens bydigesting the antigen with the endoprotease Lys-C, or by chemicallycleaving the antigen with cyanogen bromide. Peptides resulting fromeither of these procedures were separated by reversed-phase HPLC on aVydac C18 column using a mobile phase of 0.05% (v/v) trifluoroaceticacid (TFA) with a gradient of acetonitrile containing 0.05% (v/v) TFA(1%/min). The eluent was monitored at 214 nm. Major internal peptideswere identified by their UV absorbance, and their N-terminal sequenceswere determined as described above.

[0168] Using the procedures described above, six soluble M. vaccaeantigens, designated GVc-1, GVc-2, GVc-7, GVc-13, GVc-20 and GVc-22,were isolated. Determined N-terminal and internal sequences for GVc-1are shown in SEQ ID NOS: 1, 2 and 3, respectively; the N-terminalsequence for GVc-2 is shown in SEQ ID NO: 4; internal sequences forGVc-7 are shown in SEQ ID NOS: 5-8; internal sequences for GVc-13 areshown in SEQ ID NOS: 9-11; internal sequence for GVc-20 is shown in SEQID NO: 12; and N-terminal and internal sequences for GVc-22 are shown inSEQ ID NO:56-59, respectively. Each of the internal peptide sequencesprovided herein begins with an amino acid residue which is assumed toexist in this position in the polypeptide, based on the known cleavagespecificity of cyanogen bromide (Met) or Lys-C (Lys).

[0169] Three additional polypeptides, designated GVc-16, GVc-18 andGVc-21, were isolated employing a preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) purification stepin addition to the preparative isoelectric focusing procedure describedabove. Specifically, fractions comprising mixtures of polypeptides fromthe preparative isoelectric focusing purification step previouslydescribed, were purified by preparative SDS-PAGE on a 15% polyacrylamidegel. The samples were dissolved in reducing sample buffer and applied tothe gel. The separated proteins were transferred to a polyinylidenedifluoride (PVDF) membrane by electroblotting in 10 mM3-(cyclohexylamino)-1-propanesulfonic acid (CAPS) buffer pH 11containing 10% (v/v) methanol. The transferred protein bands wereidentified by staining the PVDF membrane with Coomassie blue. Regions ofthe PVDF membrane containing the most abundant polypeptide species werecut out and directly introduced into the sample cartridge of the PerkinElmer/Applied BioSystems Procise 492 protein sequencer. Proteinsequences were determined as described above. The N-terminal sequencesfor GVc-16, GVc-18 and GVc-21 are provided in SEQ ID NOS: 13, 14 and 15,respectively.

[0170] Additional antigens, designated GVc-12, GVc-14, GVc-15, GVc-17and GVc-19, were isolated employing a preparative SDS-PAGE purificationstep in addition to the chromatographic procedures described above.Specifically, fractions comprising a mixture of antigens from the VydacC4 HPLC purification step previously described were fractionated bypreparative SDS-PAGE on a polyacrylamide gel. The samples were dissolvedin non-reducing sample buffer and applied to the gel. The separatedproteins were transferred to a PVDF membrane by electroblotting in 10 mMCAPS buffer, pH 11 containing 10% (v/v) methanol. The transferredprotein bands were identified by staining the PVDF membrane withCoomassie blue. Regions of the PVDF membrane containing the mostabundant polypeptide species were cut out and directly introduced intothe sample cartridge of the Perkin Elmer/Applied BioSystems Procise 492protein sequencer. Protein sequences were determined as described above.The determined N-terminal sequences for GVc-12, GVc-14, GVc-15, GVc-17and GVc-19 are provided in SEQ ID NOS: 16-20, respectively.

[0171] All of the above amino acid sequences were compared to knownamino acid sequences in the SwissProt data base (version R32) using theGeneAssist system. No significant homologies to the amino acid sequencesGVc-2 to GVc-22 were obtained. The amino acid sequence for GVc-1 wasfound to bear some similarity to sequences previously identified from M.bovis and M. tuberculosis. In particular, GVc-1 was found to have somehomology with M. tuberculosis MPT83, a cell surface protein, as well asMPT70. These proteins form part of a protein family (Harboe et al.,Scand. J. Immunol. 42:46-51, 1995).

[0172] Subsequent studies led to the isolation of DNA sequences forGVc-13, GVc-14 and GVc-22 (SEQ ID NO: 142, 107 and 108, respectively).The corresponding predicted amino acid sequences for GVc-13, GVc-14 andGVc-22 are provided in SEQ ID NO: 143, 109 and 110, respectively.Further studies with GVc-22 suggested that only a part of the geneencoding GVc-22 was cloned. When sub-cloned into the expression vectorpET16, no protein expression was obtained. Subsequent screening of theM. vaccae BamHI genomic DNA library with the incomplete gene fragmentled to the isolation of the complete gene encoding GVc-22. Todistinguish between the full-length clone and the partial GVc-22, theantigen expressed by the full-length gene was called GV-22B. Thedetermined nucleotide sequence of the gene encoding GV-22B and thepredicted amino acid sequence are provided in SEQ ID NOS: 144 and 145respectively.

[0173] Amplifications primers AD86 and AD112 (SEQ ID NO: 60 and 61,respectively) were designed from the amino acid sequence of GVc-1 (SEQID NO: 1) and the M. tuberculosis MPT70 gene sequence. Using theseprimers, a 310 bp fragment was amplified from M. vaccae genomic DNA andcloned into EcoRV-digested vector pBluescript II SK⁺ (Stratagene). Thesequence of the cloned insert is provided in SEQ ID NO: 62. The insertof this clone was used to screen a M. vaccae genomic DNA libraryconstructed in lambda ZAP-Express (Stratgene, La Jolla, Calif.). Theclone isolated contained an open reading frame with homology to the M.tuberculosis antigen MPT83 and was re-named GV-1/83. This gene also hadhomology to the M. bovis antigen MPB83. The determined nucleotidesequence and predicted amino acid sequences are provided in SEQ ID NOS:146 and 147 respectively.

[0174] From the amino acid sequences provided in SEQ ID NOS: 1 and 2,degenerate oligonucleotides EV59 and EV61 (SEQ ID NOS: 148 and 149respectively) were designed. Using PCR, a 100 bp fragment was amplified,cloned into plasmid pBluescript II SK⁺ and sequenced (SEQ ID NO: 150)following standard procedures (Sambrook et al., Ibid ) The cloned insertwas used to screen a M. vaccae genomic DNA library constructed in lambdaZAP-Express. The clone isolated had homology to M. tuberculosis antigenMPT70 and M. bovis antigen MPB70, and was named GV-1/70. The determinednucleotide sequence and predicted amino acid sequence for GV-1/70 areprovided in SEQ ID NOS: 151 and 152, respectively. For expression andpurification, the genes encoding GV1/83, GV1/70, GVc-13, GVc-14 andGV-22B were sub-cloned into the expression vector pET16 (Novagen,Madison, Wis.). Expression and purification were carried out accordingto the manufacturer's protocol.

[0175] The purified polypeptides were screened for the ability to induceT-cell proliferation and IFN-γ in peripheral blood cells from immunehuman donors. These donors were known to be PPD (purified proteinderivative from M. tuberculosis) skin test positive and their T cellswere shown to proliferate in response to PPD. Donor PBMCs and crudesoluble proteins from M. vaccae culture filtrate were cultured in mediumcomprising RPMI 1640 supplemented with 10% (v/v) autologous serum,penicillin (60 mg/ml), streptomycin (100 mg/ml), and glutamine (2 mM).

[0176] After 3 days, 50 μl of medium was removed from each well for thedetermination of IFN-γ levels, as described below. The plates werecultured for a further 4 days and then pulsed with 1 mCi/well oftritiated thymidine for a further 18 hours, harvested and tritium uptakedetermined using a scintillation counter. Fractions that stimulatedproliferation in both replicates two-fold greater than the proliferationobserved in cells cultured in medium alone were considered positive.

[0177] IFN-γ was measured using an enzyme-linked immunosorbent assay(ELISA). ELISA plates were coated with a mouse monoclonal antibodydirected to human IFN-g (Endogen, Wobural, Mass.) 1 mg/mlphosphate-buffered saline (PBS) for 4 hours at 4° C. Wells were blockedwith PBS containing 0.2% Tween 20 for 1 hour at room temperature. Theplates were then washed four times in PBS/0.2% Tween 20, and samplesdiluted 1:2 in culture medium in the ELISA plates were incubatedovernight at room temperature. The plates were again washed, and abiotinylated polyclonal rabbit anti-human IFN-γ serum (Endogen), dilutedto 1 mg/ml in PBS, was added to each well. The plates were thenincubated for 1 hour at room temperature, washed, and horseradishperoxidase-coupled avidin A (Vector Laboratories, Burlingame, Calif.)was added at a 1:4,000 dilution in PBS. After a further 1 hourincubation at room temperature, the plates were washed andorthophenylenediamine (OPD) substrate added. The reaction was stoppedafter 10 min with 10% (v/v) HCl. The optical density (OD) was determinedat 490 nm. Fractions that resulted in both replicates giving an ODtwo-fold greater than the mean OD from cells cultured in medium alonewere considered positive.

[0178] Examples of polypeptides containing sequences that stimulateperipheral blood mononuclear cells (PBMC) T cells to proliferate andproduce IFN-γ are shown in Table 9, wherein (−) indicates a lack ofactivity, (+/−) indicates polypeptides having a result less than twicehigher than background activity of control media, (+) indicatespolypeptides having activity two to four times above background, and(++) indicates polypeptides having activity greater than four timesabove background. TABLE 9 Examples of Polypeptides Stimulating HumanPeripheral Blood Mononuclear Cells Antigen Proliferation IFN-γ GVc-1 +++/− GVc-2 + ++ GVc-7 +/− − GVc-13 + ++ GVc-14 ++ + GVc-15 + + GVc-20 + +

EXAMPLE 6 Purification and Characterisation of Polypeptides from M.vaccae Culture Filtrate by 2-Dimensional Polyacrylamide GelElectrophoresis

[0179]M. vaccae soluble proteins were isolated from culture filtrateusing 2-dimensional polyacrylamide gel electrophoresis as describedbelow. Unless otherwise noted, all percentages in the following exampleare weight per volume.

[0180]M. vaccae (ATCC Number 15483) was cultured in sterile Medium 90 at37° C. M. tuberculosis strain H37Rv (ATCC number 27294) was cultured insterile Middlebrook 7H9 medium with Tween 80 and oleicacid/albumin/dextrose/catalase additive (Difco Laboratories, Detroit,Mich.). The cells were harvested by centrifugation, and transferred intosterile Middlebrook 7H9 medium with glucose at 37° C. for one day. Themedium was then centrifuged (leaving the bulk of the cells) and filteredthrough a 0.45 μm filter into sterile bottles. The culture filtrate wasconcentrated by lyophilisation, and re-dissolved in MilliQ water. Asmall amount of insoluble material was removed by filtration through a0.45 μm membrane filter.

[0181] The culture filtrate was desalted by membrane filtration in a 400ml Amicon stirred cell which contained a 3,000 Da MWCO membrane. Thepressure was maintained at 60 psi using nitrogen gas. The culturefiltrate was repeatedly concentrated by membrane filtration and dilutedwith water until the conductivity of the sample was less than 1.0 mS.This procedure reduced the 20 l volume to approximately 50 ml. Proteinconcentrations were determined by the Bradford protein assay (Bio-Rad,Hercules, Calif., USA).

[0182] The desalted culture filtrate was fractionated by ion exchangechromatography on a column of Q-Sepharose (Pharmacia Biotech) (16×100mm) equilibrated with 10 mM TrisHCl buffer pH 8.0. Polypeptides wereeluted with a linear gradient of NaCl from 0 to 1.0 M in the abovebuffer system. The column eluent was monitored at a wavelength of 280nm.

[0183] The pool of polypeptides eluting from the ion exchange columnwere fractionated by preparative 2-D gel electrophoresis. Samplescontaining 200-500 μg of polypeptide were made 8M in urea and applied topolyacrylamide isoelectric focusing rod gels (diameter 2 mm, length 150mm, pH 5-7). After the isoelectric focusing step, the first dimensiongels were equilibrated with reducing buffer and applied to seconddimension gels (16% polyacrylamide). FIGS. 4A and 4B are the 2-D gelpatterns observed with M. vaccae culture filtrate and M. tuberculosisH37Rv culture filtrate, respectively. Polypeptides from the seconddimension separation were transferred to PVDF membranes byelectroblotting in 10 mM CAPS buffer pH 11 containing 10% (v/v)methanol. The PVDF membranes were stained for protein with Coomassieblue. Regions of PVDF containing polypeptides of interest were cut outand directly introduced into the sample cartridge of the PerkinElmer/Applied BioSystems Procise 492 protein sequencer. The polypeptideswere sequenced from the amino terminal end using traditional Edmanchemistry. The amino acid sequence was determined for each polypeptideby comparing the retention time of the PTH amino acid derivative to theappropriate PTH derivative standards. Using these procedures, elevenpolypeptides, designated GVs-1, GVs-3, GVs-4, GVs-5, GVs-6, GVs-8,GVs-9, GVs-10, GVs-11, GV-34 and GV-35 were isolated. The determinedN-terminal sequences for these polypeptides are shown in SEQ ID NOS:21-29, 63 and 64, respectively. Using the purification proceduredescribed above, more protein was purified to extend the amino acidsequence previously obtained for GVs-9. The extended amino acid sequencefor GVs-9 is provided in SEQ ID NO:65. Further studies resulted in theisolation of the DNA sequences for GVs-9 (SEQ ID NO: 111) and GV-35 (SEQID NO: 155). The corresponding predicted amino acid sequences areprovided in SEQ ID NO: 112 and 156, respectively. An extended DNAsequence for GVs-9 is provided in SEQ ID NO: 153, with the correspondingpredicted amino acid sequence being provided in SEQ ID NO: 154.

[0184] All of these amino acid sequences were compared to known aminoacid sequences in the SwissProt data base (version R32) using theGeneAssist system. No significant homologies were obtained, with theexceptions of GVs-3, GVs-4, GVs-5 and GVs-9. GVs-9 was found to bearsome homology to two previously identified M. tuberculosis proteins,namely M. tuberculosis cutinase precursor and a M. tuberculosishypothetical 22.6 kDa protein. GVs-3, GVs-4 and GVs-5 were found to bearsome similarity to the antigen 85A and 85B proteins from M. leprae (SEQID NOS: 30 and 31, respectively), M. tuberculosis (SEQ ID NOS: 32 and33, respectively) and M. bovis (SEQ ID NOS: 34 and 35, respectively),and the antigen 85C proteins from M. leprae (SEQ ID NO: 36) and M.tuberculosis (SEQ ID NO: 37).

EXAMPLE 7 DNA Cloning Strategy for the M. vaccae Antigen 85 Series

[0185] Probes for antigens 85A, 85B, and 85C were prepared by thepolymerase chain reaction (PCR) using degenerate oligonucleotides (SEQID NOS: 38 and 39) designed to regions of antigen 85 genomic sequencethat are conserved between family members in a given mycobacterialspecies, and between mycobacterial species. These oligonucleotides wereused under reduced stringency conditions to amplify target sequencesfrom M. vaccae genomic DNA. An appropriately-sized 485 bp band wasidentified, purified, and cloned pBluescript II SK⁺ (Stratagene, LaJolla, Calif.). Twenty-four individual colonies were screened at randomfor the presence of the antigen 85 PCR product, then sequenced using thePerkin Elmer/Applied Biosystems Model 377 automated sequencer and theM13-based primers, T3 and T7. Homology searches of the GenBank databasesshowed that twenty-three clones contained insert with significanthomology to published antigen 85 genes from M. tuberculosis and M.bovis. Approximately half were most homologous to antigen 85C genesequences, with the remainder being more similar to antigen 85Bsequences. In addition, these two putative M. vaccae antigen 85 genomicsequences were 80% homologous to one another. Because of this highsimilarity, the antigen 85C PCR fragment was chosen to screen M. vaccaegenomic libraries at low stringency for all three antigen 85 genes.

[0186] An M. vaccae genomic library was created in lambda Zap-Express(Stratagene, La Jolla, Calif.) by cloning BamHI partially-digested M.vaccae genomic DNA into similarly-digested vector, with 3.4×10⁵independent plaque-forming units resulting. For screening purposes,twenty-seven thousand plaques from this non-amplified library wereplated at low density onto eight 100 cm² plates. For each plate,duplicate plaque lifts were taken onto Hybond-N⁺ nylon membrane(Amersham International, United Kingdom), and hybridised underreduced-stringency conditions (55° C.) to the radiolabelled antigen 85CPCR product. Autoradiography demonstrated that seventy-nine plaquesconsistently hybridised to the antigen 85C probe under these conditions.Thirteen positively-hybridising plaques were selected at random forfurther analysis and removed from the library plates, with each positiveclone being used to generate secondary screening plates containing abouttwo hundred plaques. Duplicate lifts of each plate were taken usingHybond-N nylon membrane, and hybridised under the conditions used inprimary screening. Multiple positively-hybridising plaques wereidentified on each of the thirteen plates screened. Two well-isolatedpositive phage from each secondary plate were picked for furtheranalysis. Using in vitro excision, twenty-six plaques were convertedinto phagemid, and restriction-mapped. It was possible to group clonesinto four classes on the basis of this mapping. Sequence data from the5′ and 3′ ends of inserts from several representatives of each group wasobtained using the Perkin Elmer/Applied Biosystems Division Model 377automated sequencer and the T3 and T7 primers. Sequence homologies weredetermined using FASTA analysis of the GenBank databases with theGeneAssist software package. Two of these sets of clones were found tobe homologous to M. bovis and M. tuberculosis antigen 85A genes, eachcontaining either the 5′ or 3′ ends of the M. vaccae gene (this gene wascleaved during library construction as it contains an internal BamHIsite). The remaining clones were found to contain sequences homologousto antigens 85B and 85C from a number of mycobacterial species. Todetermine the remaining nucleotide sequence for each gene, appropriatesubclones were constructed and sequenced. Overlapping sequences werealigned using the DNA Strider software. The determined DNA sequences forM. vaccae antigens 85A, 85B and 85C are shown in SEQ ID NOS: 40-42,respectively, with the predicted amino acid sequences being shown in SEQID NOS: 43-45, respectively.

[0187] The M. vaccae antigens GVs-3 and GVs-5 were expressed andpurified as follows. Amplification primers were designed from the insertsequences of GVs-3 and GVs-5 (SEQ ID NO: 40 and 42, respectively) usingsequence data downstream from the putative leader sequence and the 3′end of the clone. The sequences of the primers for GVs-3 are provided inSEQ ID NO: 66 and 67, and the sequences of the primers for GVs-5 areprovided in SEQ ID NO: 68 and 69. A XhoI restriction site was added tothe primers for GVs-3, and EcoRI and BamHI restriction sites were addedto the primers for GVs-5 for cloning convenience. Followingamplification from genomic M. vaccae DNA, fragments were cloned into theappropriate site of pProEX HT prokaryotic expression vector (Gibco BRL,Life Technologies, Gaithersburg, Md.) and submitted for sequencing toconfirm the correct reading frame and orientation. Expression andpurification of the recombinant protein was performed according to themanufacturer's protocol.

[0188] Expression of a fragment of the M. vaccae antigen GVs-4 (antigen85B homolog) was performed as follows. The primers AD58 and AD59,described above, were used to amplify a 485 bp fragment from M. vaccaegenomic DNA. This fragment was gel-purified using standard techniquesand cloned into EcoRV-digested pBluescript. The base sequences ofinserts from five clones were determined and found to be identical toeach other. These inserts had highest homology to Ag85B from M.tuberculosis. The insert from one of the clones was subcloned into theEcoRI/XhoI sites of pProEX HT prokaryotic expression vector (Gibco BRL),expressed and purified according to the manufacturer's protocol. Thisclone was renamed GV-4P because only a part of the gene was expressed.The amino acid and DNA sequences for the partial clone GV-4P areprovided in SEQ ID NO: 70 and 106, respectively.

[0189] Similar to the cloning of GV-4P, the amplification primers AD58and AD59 were used to amplify a 485 bp fragment from a clone containingGVs-5 (SEQ ID NO:42). This fragment was cloned into the expressionvector pET16 and was called GV-5P. The determined nucleotide sequenceand predicted amino acid sequence of GV-5P are provided in SEQ ID NOS:157 and 158, respectively.

[0190] The ability of purified recombinant GVs-3, GV-4P and GVs-5 tostimulate proliferation of T cells and interferon-y production in humanPBL was assayed as described above in Example 4. The results of thisassay are shown in Table 10, wherein (−) indicates a lack of activity,(+/−) indicates polypeptides having a result less than twice higher thanbackground activity of control media, (+) indicates polypeptides havingactivity two to four times above background, (++) indicates polypeptideshaving activity greater than four times above background, and NDindicates not determined. TABLE 10 Donor Donor Donor Donor Donor DonorG97005 G97006 G97007 G97008 G97009 G97010 Prolif IFN-γ Prolif IFN-γProlif IFN-γ Prolif IFN-γ Prolif IFN-γ Prolif IFN-γ GVs-3 ++ + ND ND ++++ ++ ++ ++ +/− + ++ GV- + +/− ND ND + ++ ++ ++ +/− +/− +/− ++ 4P GVs-5++ ++ ++ ++ ++ ++ + ++ ++ + + ++

EXAMPLE 8 DNA Cloning Strategy for M. vaccae Antigens

[0191] An 84 bp probe for the M. vaccae antigen GVc-7 was amplifiedusing degenerate oligonucleotides designed to the determined amino acidsequence of GVc-7 (SEQ ID NOS: 5-8). This probe was used to screen a M.vaccae genomic DNA library as described in Example 4. The determinednucleotide sequence for GVc-7 is shown in SEQ ID NO: 46 and predictedamino acid sequence in SEQ ID NO: 47. Comparison of these sequences withthose in the databank revealed homology to a hypothetical 15.8 kDamembrane protein of M. tuberculosis.

[0192] The sequence of SEQ ID NO: 46 was used to design amplificationprimers (provided in SEQ ID NO: 71 and 72) for expression cloning of theGVc-7 gene using sequence data downstream from the putative leadersequence. A XhoI restriction site was added to the primers for cloningconvenience. Following amplification from genomic M. vaccae DNA,fragments were cloned into the XhoI-site of pProEX HT prokaryoticexpression vector (Gibco BRL) and submitted for sequencing to confirmthe correct reading frame and orientation. Expression and purificationof the fusion protein was performed according to the manufacturer'sprotocol.

[0193] The ability of purified recombinant GVc-7 to stimulateproliferation of T-cells and stimulation of interferon-γ production inhuman PBL was assayed as described previously in Example 4. The resultsare shown in Table 11, wherein (−) indicates a lack of activity, (+/−)indicates polypeptides having a result less than twice higher thanbackground activity of control media, indicates polypeptides havingactivity two to four times above background, and (++) indicatespolypeptides having activity greater than four times above background.TABLE 11 Donor Proliferation Interferon-γ G97005 ++ +/− G97008 ++ +G97009 + +/− G97010 +/− ++

[0194] A redundant oligonucleotide probe SEQ ID NO 73, referred to asMPG15) was designed to the GVs-8 peptide sequence shown in SEQ ID NO: 26and used to screen a M. vaccae genomic DNA library using standardprotocols.

[0195] A genomic clone containing genes encoding four different antigenswas isolated. The determined DNA sequences for GVs-8A (re-named GV-30),GVs-8B (re-named GV-31), GVs-8C (re-named GV-32) and GVs-8D (re-namedGV-33) are shown in SEQ ID NOS: 48-51, respectively, with thecorresponding amino acid sequences being shown in SEQ ID NOS: 52-55,respectively. GV-30 contains regions showing some similarity to knownprokaryotic valyl-tRNA synthetases; GV-31 shows some similarity to M.smegmatis aspartate semialdehyde dehydrogenase; and GV-32 shows somesimilarity to the H. influenza folylpolyglutamate synthase gene.GV-33contains an open reading frame which shows some similarity tosequences previously identified in M. tuberculosis and M. leprae, butwhose function has not been identified.

[0196] The determined partial DNA sequence for GV-33 is provided in SEQID NO:74 with the corresponding predicted amino acid sequence beingprovided in SEQ ID NO:75. Sequence data from the 3′ end of the cloneshowed homology to a previously identified 40.6 kDa outer membraneprotein of M. tuberculosis. Subsequent studies led to the isolation ofthe full-length DNA sequence for GV-33 (SEQ ID NO: 193). Thecorresponding predicted amino acid sequence is provided in SEQ ID NO:194.

[0197] The gene encoding GV-33 was amplified from M. vaccae genomic DNAwith primers based on the determined nucleotide sequence. This DNAfragment was cloned into EcoRv-digested pBluescript II SK⁺ (Stratagene),and then transferred to pET16 expression vector. Recombinant protein waspurified following the manufacturer's protocol.

[0198] The ability of purified recombinant GV-33 to stimulateproliferation of T-cells and stimulation of interferon-γ production inhuman PBL was assayed as described previously in Example 5. The resultsare shown in Table 12, wherein (−) indicates a lack of activity, (+/−)indicates polypeptides having a result less than twice higher thanbackground activity of control media, (+) indicates polypeptides havingactivity two to four times above background, and (++) indicatespolypeptides having activity greater than four times above background.TABLE 12 Stimulatory Activity of Polypeptides Donor ProliferationInterferon-γ G97005 ++ + G97006 ++ ++ G97007 − +/− G97008 +/− − G97009+/− − G97010 +/− ++

EXAMPLE 9 DNA Cloning Strategy for the M. vaccae Antigens GV-23. GV-24GV-25 GV-26, GV-38A and GV-38B

[0199]M. vaccae (ATCC Number 15483) was grown in sterile Medium 90 at37° C. for 4 days and harvested by centrifugation. Cells wereresuspended in 1 ml TRIzol (Gibco BRL, Life Technologies, Gaithersburg,Md.) and RNA extracted according to the standard manufacturer'sprotocol. M. tuberculosis strain H37Rv (ATCC Number 27294) was grown insterile Middlebrooke 7H9 medium with Tween 80™ and oleicacid/albumin/dextrose/catalase additive (Difco Laboratories, Detroit,Mich.) at 37° C. and harvested under appropriate laboratory safetyconditions. Cells were resuspended in 1 ml TRIzol (Gibco BRL) and RNAextracted according to the manufacturer's standard protocol.

[0200] Total M. tuberculosis and M. vaccae RNA was depleted of 16S and23S ribosomal RNA (rRNA) by hybridisation of the total RNA fraction tooligonucleotides AD10 and AD11 (SEQ ID NO: 81 and 82) complementary toM. tuberculosis rRNA. These oligonucleotides were designed frommycobacterial 16S rRNA sequences published by Bottger (FEMS Microbiol.Lett. 65:171-176, 1989) and from sequences deposited in the databanks.Depletion was done by hybridisation of total RNA to oligonucleotidesAD10 and AD11 immobilised on nylon membranes (Hybond N, AmershamInternational, United Kingdom). Hybridisation was repeated until rRNAbands were not visible on ethidium bromide-stained agarose gels. Anoligonucleotide, AD12 (SEQ ID NO: 83), consisting of 20 dATP-residues,was ligated to the 3′ ends of the enriched mRNA fraction using RNAligase. First strand cDNA synthesis was performed following standardprotocols, using oligonucleotide AD7 (SEQ ID NO: 84) containing apoly(dT) sequence.

[0201] The M. tuberculosis and M. vaccae cDNA was used as template forsingle-sided-specific PCR (3S-PCR). For this protocol, a degenerateoligonucleotide AD1 (SEQ ID NO:85) was designed based on conservedleader sequences and membrane protein sequences. After 30 cycles ofamplification using primer AD1 as 5′-primer and AD7 as 3′-primer,products were separated on a urea/polyacrylamide gel. DNA bands uniqueto M. vaccae were excised and re-amplified using primers AD1 and AD7.After gel purification, bands were cloned into pGEM-T (Promega) and thebase sequence determined.

[0202] Searches with the determined nucleotide and predicted amino acidsequences of band 12B21 (SEQ ID NOS: 86 and 87, respectively) showedhomology to the pota gene of Escherichia coli encoding the ATP-bindingprotein of the spermidine/putrescine ABC transporter complex publishedby Furuchi et al. (J. Biol. Chem. 266:20928-20933, 1991). Thespermidine/putrescine transporter complex of E. coli consists of fourgenes and is a member of the ABC transporter family. The ABC(ATP-binding Cassette) transporters typically consist of four genes: anATP-binding gene, a periplasmic, or substrate binding, gene and twotransmembrane genes. The transmembrane genes encode proteins eachcharacteristically having six membrane-spanning regions. Homologues (bysimilarity) of this ABC transporter have been identified in the genomesof Haemophilus influenza (Fleischmann et al. Science 269 :496-512, 1995)and Mycoplasma genitalium (Fraser, et al. Science, 270:397-403, 1995).

[0203] A M. vaccae genomic DNA library constructed in BamHI-digestedlambda ZAP Express (Stratagene) was probed with the radiolabelled 238 bpband 12B21 following standard protocols. A plaque was purified to purityby repetitive screening and a phagemid containing a 4.5 kb insert wasidentified by Southern blotting and hybridisation. The nucleotidesequence of the full-length M. vaccae homologue of pota (ATP-bindingprotein) was identified by subcloning of the 4.5 kb fragment and basesequencing. The gene consisted of 1449 bp including an untranslated 5′region of 320 bp containing putative −10 and −35 promoter elements. Thenucleotide and predicted amino acid sequences of the M. vaccae potahomologue are provided in SEQ ID NOS: 88 and 89, respectively.

[0204] The nucleotide sequence of the M. vaccae pota gene was used todesign primers EV24 and EV25 (SEQ ID NO: 90 and 91) for expressioncloning. The amplified DNA fragment was cloned into pProEX HTprokaryotic expression system (Gibco BRL) and expression in anappropriate E. coli host was induced by addition of 0.6 mMisopropylthio-β-galactoside (IPTG). The recombinant protein was namedGV-23 and purified from inclusion bodies according to the manufacturer'sprotocol.

[0205] A 322 bp Sal1-BamH1 subclone at the 3′-end of the 4.5 kb insertdescribed above showed homology to the potd gene, (periplasmic protein),of the spermidine/putrescine ABC transporter complex of E. coli. Thenucleotide sequence of this subclone is shown in SEQ ID NO:92. Toidentify the gene, the radiolabelled insert of this subclone was used toprobe an M. vaccae genomic DNA library constructed in the Sal1-site oflambda Zap-Express (Stratagene) following standard protocols. A clonewas identified of which 1342 bp showed homology with the potd gene of E.coli. The potd homologue of M. vaccae was identified by sub-cloning andbase sequencing. The determined nucleotide and predicted amino acidsequences are shown in SEQ ID NO: 93 and 94.

[0206] For expression cloning, primers EV26 and EV27 (SEQ ID NOS:95-96)were designed from the determined M. vaccae potd homologue. Theamplified fragment was cloned into pProEX HT Prokaryotic expressionsystem (Gibco BRL). Expression in an appropriate E. coli host wasinduced by addition of 0.6 mM IPTG and the recombinant protein namedGV-24. The recombinant antigen was purified from inclusion bodiesaccording to the protocol of the supplier.

[0207] To improve the solubility of the purified recombinant antigen,the gene encoding GV-24, but excluding the signal peptide, was re-clonedinto the expression vector, employing. amplification primers EV101 andEV102 (SEQ ID NOS: 167 and 168). The construct was designated GV-24B.The nucleotide sequence of GV-24B is provided in SEQ ID NO: 169 and thepredicted amino acid sequence in SEQ ID NO: 170. This fragment wascloned into pET16 for expression and purification of GV-24B according tothe manufacturer's protocols.

[0208] The ability of purified recombinant protein GV-23 and GV-24 tostimulate proliferation of T cells and interferon-production in humanPBL was determined as described in Example 4. The results of theseassays are provided in Table 13, wherein (−) indicates a lack ofactivity, (+/−) indicates polypeptides having a result less than twicehigher than background activity of control media, (+) indicatespolypeptides having activity two to four times above background, (++)indicates polypeptides having activity greater than four times abovebackground, and (ND) indicates not determined. TABLE 13 Donor DonorDonor Donor Donor Donor G97005 G97006 G97007 G97008 G97009 G97010 ProlifIFN-γ Prolif IFN-γ Prolif IFN-γ Prolif IFN-γ Prolif IFN-γ Prolif IFN-γGV-23 ++ ++ ++ ++ + + ++ ++ + − + ++ GV-24 ++ + ++ + ND ND + +/− + +/−+/− ++

[0209] Base sequence adjacent to the M. vaccae potd gene-homologue wasfound to show homology to the potb gene of the spermidine/putrescine ABCtransporter complex of E.coli, which is one of two transmembraneproteins in the ABC transporter complex. The M. vaccae potb homologue(referred to as GV-25) was identified through further subcloning andbase sequencing. The determined nucleotide and predicted amino acidsequences for GV-25 are shown in SEQ ID NOS: 97 and 98, respectively.

[0210] Further subcloning and base sequence analysis of the adjacent 509bp failed to reveal significant homology to PotC, the secondtransmembrane protein of E.coli, and suggests that a secondtransmembrane protein is absent in the M. vaccae homologue of the ABCtransporter. An open reading frame with homology to M. tuberculosisacetyl-CoA acetyl transferase, however, was identified starting 530 bpdownstream of the transmembrane protein and the translated protein wasnamed GV-26. The determined partial nucleotide sequence and predictedamino acid sequence for GV-26 are shown in SEQ ID NO:99 and 100.

[0211] Using a protocol similar to that described above for theisolation of GV-23, the 3S-PCR band 12B28 (SEQ ID NO: 119) was used toscreen the M. vaccae genomic library constructed in the BamHI-site oflambda ZAP-Express (Stratagene). The clone isolated from the librarycontained a novel open reading frame and the antigen encoded by thisgene was named GV-38A. The determined nucleotide sequence and predictedamino acid sequence of GV-38A are shown in SEQ ID NO: 120 and 121,respectively. Subsequent studies led to the isolation of an extended DNAsequence for GV-38A, provided in SEQ ID NO: 171. The corresponding aminoacid sequence is provided in SEQ ID NO: 172. Comparison of thesesequences with those in the database revealed only a limited amount ofhomology to an unknown M. tuberculosis protein previously identified incosmid MTCY428.12.

[0212] Upstream of the GV-38A gene, a second novel open reading framewas identified and the antigen encoded by this gene was named GV-38B.The determined 5′ and 3′ nucleotide sequences for GV-38B are provided inSEQ ID NO: 122 and 123, respectively, with the corresponding predictedamino acid sequences being provided in SEQ ID NO: 124 and 125,respectively. Further studies led to the isolation of the full-lengthDNA sequence for GV-38B, provided in SEQ ID NO: 173. The correspondingamino acid sequence is provided in SEQ ID NO: 174. This protein wasfound to show only a limited amount of homology to an unknown M.tuberculosis protein identified as a putative open reading frame incosmid MTCY428.11 (SPTREMBL: P71914).

[0213] Both the GV-38A and GV-38B antigens were amplified for expressioncloning into pET16 (Novagen). GV-38A was amplified with primers KR11 andKR12 (SEQ ID NO: 126 and 127) and GV-38B with primers KR13 and KR14 (SEQID NO: 128 and 129). Protein expression in the host cells BL21 (DE3) wasinduced with 1 mM IPTG, however no protein expression was obtained fromthese constructs. Hydrophobic regions were identified in the N-terminiof antigens GV-38A and GV-38B which may inhibit expression of theseconstructs. The hydrophobic region present in GV-38A was identified as apossible transmembrane motif with six membrane spanning regions. Toexpress the antigens without the hydrophobic regions, primers KR20 forGV-38A, (SEQ ID NO: 130) and KR21 for GV-38B (SEQ ID NO: 131) weredesigned. The truncated GV-38A gene was amplified with primers KR20 andKR12, and the truncated GV-38B gene with KR21 and KR14. The determinednucleotide sequences of truncated GV-38A and GV-38B are shown in SEQ IDNO: 132 and 133, respectively, with the corresponding predicted aminoacid sequences being shown in SEQ ID NO: 134 and 135, respectively.Extended DNA sequences for truncated GV-38A and GV-38B are provided inSEQ ID NO: 175 and 176, respectively, with the corresponding amino acidsequences being provided in SEQ ID NO: 177 and 178, respectively.

EXAMPLE 10 Purification and Characterisation of Polypeptides from M.vaccae Culture Filtrate by Preparative Isoelectric Focusing andPreparative Polyacrylamide Gel Electrophoresis

[0214]M. vaccae soluble proteins were isolated from culture filtrateusing preparative isoelectric focusing and preparative polyacrylamidegel electrophoresis as described below. Unless otherwise noted, allpercentages in the following example are weight per volume.

[0215]M. vaccae (ATCC Number 15483) was cultured in 250 l sterile Medium90 which had been fractionated by ultrafiltration to remove all proteinsof greater than 10 kDa molecular weight. The medium was centrifuged toremove the bacteria, and sterilised by filtration through a 0.45 μfilter. The sterile filtrate was concentrated by ultrafiltration over a10 kDa molecular weight cut-off membrane.

[0216] Proteins were isolated from the concentrated culture filtrate byprecipitation with 10% trichloroacetic acid. The precipitated proteinswere re-dissolved in 100 mM Tris.HCl pH 8.0 and re-precipitated by theaddition of an equal volume of acetone. The acetone precipitate wasdissolved in water, and proteins were re-precipitated by the addition ofan equal volume of chloroform:methanol 2:1 (v/v). The chloroformmethanol precipitate was dissolved in water, and the solution wasfreeze-dried.

[0217] The freeze-dried protein was dissolved in iso-electric focusingbuffer, containing 8 M deionised urea, 2% Triton X-100, 10 mMdithiothreitol and 2% ampholytes (pH 2.5-5.0). The sample wasfractionated by preparative iso-electric focusing on a horizontal bed ofUltrodex gel at 8 watts constant power for 16 hours. Proteins wereeluted from the gel bed fractions with water and concentrated byprecipitation with 10% trichloroacetic acid.

[0218] Pools of fractions containing proteins of interest wereidentified by analytical polyacrylamide gel electrophoresis andfractionated by preparative polyacrylamide gel electrophoresis. Sampleswere fractionated on 12.5% SDS-PAGE gels, and electroblotted ontonitrocellulose membranes. Proteins were located on the membranes bystaining with Ponceau Red, destained with water and eluted from themembranes with 40% acetonitrile/0.1M ammonium bicarbonate pH 8.9 andthen concentrated by lyophilisation.

[0219] Eluted proteins were assayed for their ability to induceproliferation and interferon-γ secretion from the peripheral bloodlymphocytes of immune donors as detailed in Example 4. Proteins inducinga strong response in these assays were selected for further study.

[0220] Selected proteins were further purified by reversed-phasechromatography on a Vydac Protein C4 column, using a trifluoroaceticacid-acetonitrile system Purified proteins were prepared for proteinsequence determination by SDS-polyacrylamide gel electrophoresis, andelectroblotted onto PVDF membranes. Protein sequences were determined asin Example 5. The proteins were named GV-40, GV-41, GV-42, GV-43 andGV-44. The determined N-terminal sequences for these polypeptides areshown in SEQ ID NOS:101-105, respectively. Subsequent studies led to theisolation of a 5′, middle fragment and 3′ DNA sequence for GV-42 (SEQ IDNO: 136, 137 and 138, respectively). The corresponding predicted aminoacid sequences are provided in SEQ ID NO: 139, 140 and 141,respectively.

[0221] Following standard DNA amplification and cloning procedures asdescribed in Example 7, the genes encoding GV-41 and GV-42 were cloned.The determined nucleotide sequences are provided in SEQ ID NOS: 179 and180, respectively, and the predicted amino acid sequences in SEQ ID NOS:181 and 182. GV-41 had homology to the ribosome recycling factor of M.tuberculosis and M. leprae, and GV-42 had homology to a M. aviumfibronectin attachment protein FAP-A. Within the full-length sequence ofGV-42, the amino acid sequence determined for GV-43 (SEQ ID NO: 104) wasidentified, indicating that the amino acid sequences for GV-42 and GV-43were obtained from the same protein.

[0222] Murine polyclonal antisera were prepared against GV-40 and GV-44following standard procedures. These antisera were used to screen a M.vaccae genomic DNA library consisting of randomly sheared DNA fragments.Clones encoding GV-40 and GV-44 were identified and sequenced. Thedetermined nucleotide sequence of the partial gene encoding GV-40 isprovided in SEQ ID NO: 183 and the predicted amino acid sequence in SEQID NO: 184. The nucleotide sequence of the gene encoding GV-44 isprovided in SEQ ID NO: 185, and the predicted amino acid sequence in SEQID NO: 186. Homology of GV-40 to M. leprae Elongation factor G wasfound. GV-44 had homology to M. leprae glyceraldehyde-3-phosphatedehydrogenase.

EXAMPLE 11 DNA Cloning Strategy for the DD-M. vaccae Antigen GV-45

[0223] Proteins were extracted from DD-M. vaccae (500 mg; prepared asdescribed in Example 1) by suspension in 10 ml 2% SDS/PBS and heating to50° C. for 2 h. The insoluble residue was removed by centrifugation, andproteins precipitated from the supernatant by adding an equal volume ofacetone and incubating at −20° C. for 1 hr. The precipitated proteinswere collected by centrifugation, dissolved in reducing sample buffer,and fractionated by preparative SDS-polyacrylamide gel electrophoresis.The separated proteins were electroblotted onto PVDF membrane in 10 mMCAPS/0.01% SDS pH 11.0, and N-terminal sequences were determined in agas-phase sequenator.

[0224] The amino acid sequence obtained from these experiments wasdesignated GV-45. The determined N-terminal sequence for GV-45 isprovided in SEQ ID NO: 187.

[0225] From the amino acid sequence of GV-45, degenerateoligonucleotides KR32 and KR33 (SEQ ID NOS: 188 and 189, respectively)were designed. A 100 bp fragment was amplified, cloned into plasmidpBluescript II SK⁺ (Stratagene, La Jolla, Calif.) and sequenced (SEQ IDNO: 190) following standard procedures (Sambrook et al., Ibid ). Thecloned insert was used to screen a M. vaccae genomic DNA libraryconstructed in the BamHI-site of lambda ZAP-Express (Stratagene). Theisolated clone showed homology to a 35 kDa M. tuberculosis and a 22 kDaM. leprae protein containing bacterial histone-like motifs at theN-terminus and a unique C-terminus consisting of a five amino acid basicrepeat. The determined nucleotide sequence for GV-45 is provided in SEQID NO: 191, with the corresponding predicted amino acid sequence beingprovided in SEQ ID NO: 192.

Example 12 Effect of Immunisation with M. vaccae on Immune SystemDisorders in Mice

[0226] This example illustrates that both heat-killed M. vaccae andDD-M. vaccae, when administered to mice via the intranasal route, areable to inhibit the development of an allergic immune response in thelungs and to suppress Th2 immune responses. Such responses are believedto play a role in skin disorders such as atopic dermatitis and allergiccontact dermatitis. The ability of heat-killed M. vaccae and DD-M.vaccae to inhibit the development of allergic immune responses wasdemonstrated in a mouse model of the asthma-like allergen specific lungdisease. The severity of this allergic disease is reflected in the largenumbers of eosinophils that accumulate in the lungs.

[0227] C57BL/6J mice were given 2 μg ovalbumin in 100 μl alum (Aluminiumhydroxide) adjuvant by the intraperitoneal route at time 0 and 14 days,and subsequently given 100 μg ovalbumin in 50 μl phosphate bufferedsaline (PBS) by the intranasal route on day 28. The mice accumulatedeosinophils in their lungs as detected by washing the airways of theanaesthetised mice with saline, collecting the washings (broncheolarlavage or BAL), and counting the numbers of eosinophils.

[0228] As shown in FIGS. 4A and B, groups of seven mice administeredeither 10 or 1000 μg of heat-killed M. vaccae (FIG. 4A), or 10, 100 or200 μg of DD-M. vaccae (FIG. 4B) intranasally 4 weeks before intranasalchallenge with ovalbumin, had reduced percentages of eosinophils in theBAL cells collected 5 days after challenge with ovalbumin compared tocontrol mice. Control mice were given intranasal PBS. Live M. bovis BCGat a dose of 2×10⁵ colony forming units also reduced lung eosinophilia.The data in FIGS. 4A and B show the mean and SEM per group of mice.

[0229]FIGS. 4C and D show that mice given either 1000 μg of heat-killedM. vaccae (FIG. 4C) or 200 μg of DD-M. vaccae (FIG. 4D) intranasally aslate as one week before challenge with ovalbumin had reduced percentagesof eosinophils compared to control mice. In contrast, treatment withlive BCG one week before challenge with ovalbumin did not inhibit thedevelopment of lung eosinophilia when compared with control mice.

[0230] As shown in FIG. 4E, immunisation with either 1 mg of heat-killedM. vaccae or 200 μg of DD-M. vaccae, given either intranasally (i.n.) orsubcutaneously (s.c.), reduced lung eosinophilia following challengewith ovalbumin when compared to control animals given PBS. In the sameexperiment, immunization with BCG of the Pasteur (BCG-P) and Connought(BCG-C) strains prior to challenge with ovalbumin also reduced thepercentage of eosinophils in the BAL of mice.

[0231] Eosinophils are blood cells that are prominent in the airways inallergic asthma. The secreted products of eosinophils contribute to theswelling and inflammation of the mucosal linings of the airways inallergic asthma. The data shown in FIGS. 4A-E indicate that treatmentwith heat-killed M. vaccae or DD-M. vaccae reduces the accumulation oflung eosinophils, and may be useful in reducing inflammation associatedwith eosinophilia in the airways, nasal mucosal and upper respiratorytract. Administration of heat-killed M. vaccae or DD-M. vaccae maytherefore reduce the severity of asthma and other diseases that involvesimilar immune abnormalities, such as allergic rhinitis and certainallergic skin disorders.

[0232] In addition, serum samples were collected from mice in theexperiment described in FIG. 4E and the level of antibodies to ovalbuminwas measured by standard enzyme-linked immunoassay (EIA). As shown inTable 14 below, sera from mice infected with BCG had higher levels ofovalbumin specific IgG1 than sera from PBS controls. In contrast, miceimmunized with M. vaccae or DD-M. vaccae had similar or lower levels ofovalbumin-specific IgG1. As IgG1 antibodies are characteristic of a Th2immune response, these results are consistent with the suppressiveeffects of heat-killed M. vaccae and DD-M. vaccae on the asthma-inducingTh2 immune responses, and indicate that heat-killed M. vaccae and DD-M.vaccae may be usefully employed to suppress Th2 immune responses in skindisorders such as atopic dermatitis, allergic contact dermatitis andalopecia areata. TABLE 14 LOW ANTIGEN-SPECIFIC IgG1 SERUM LEVELS IN MICEIMMUMZED WITH HIBAT-KILLED M. VACCAE OR DD-M. VACCAE Serum IgG1Treatment Group Mean SEM M.vaccae i.n. 185.00 8.3 M. vaccae s.c. 113.648.0 DD-M. vaccae i.n. 96.00 8.1 DD-M. vaccae s.c. 110.00 4.1 BCG,Pasteur 337.00 27.2 BCG, Connaught 248.00 46.1 PBS 177.14 11.4

EXAMPLE 14 Effect of DD-M. vaccae on IL-10 Production in THP-1 Cells

[0233] Psoriasis is characterised by a pronounced T cell infiltrate thatis thought to be central in driving ongoing skin inflammation. Variousstudies have shown that these cells produce a wide variety of cytokines,such as interleukin-2 (IL-2), IFNγ and TNFα, which are known to beproduced by Th1 cells. IL-10 inhibits the cytokine production of Th1cells and plays a key role in the suppression of experimentally-inducedinflammatory responses in skin (Berg et al., J. Exp. Med., 182:99-108,1995). Recently, IL-10 has been used successfully in two clinical trialsto treat psoriatic patients (Reich et al., J. Invest. Dermatol, 111:1235-1236, 1998 and Asadullah et al., J. Clin. Invest., 101:783-794,1998). It is therefore possible that DD-M. vaccae inhibits skininflammation in psoriasis patients by stimulating the production ofIL-10. To test this hypothesis, the levels of IL-10 produced by a humanmonocytic cell line (THP-1) cultured in the presence of DD-M. vaccaewere assessed.

[0234] THP-1 cells (ATCC (Rockville, Md.), TIB-202) were cultured inRPMI medium (Gibco BRL Life Technologies) supplemented with 0.5 mg/lstreptomycin, 500 U/l penicillin, 2 mg/1 L-glutamine, 5×10⁻⁵ Mβ-mercaptoethanol and 5% fetal bovine serum (FBS). One day prior to theassay, the cells were subcultured in fresh media at 5×10⁵ cells/ml.Cells were incubated at 37° C. in humidified air containing 5% CO₂ for24 hours and then aspirated and washed by centrifugation with 50 ml ofmedia. The cells were re-suspended in 5 ml of media and the cellconcentration and viability determined by staining with Trypan blue(Sigma, St Louis Mich.) and analysis under a haemocytometer. DD-M.vaccae (prepared as described above) in 50 μl PBS and control stimulantswere added in triplicate to wells of a 96 well plate containing 100 μlof medium and appropriate dilutions were prepared. Lipopolysaccharide(LPS) (300 μg/ml; Sigma) and PBS were used as controls. To each well,100 μl of cells were added at a concentration of 2×10⁶ cells/ml and theplates incubated at 37° C. in humidified air containing 5% CO₂ for 24hours. The level of IL-10 in each well was determined using the HumanIL-10 ELISA reagents (PharMingen, San Diego Calif.) according to themanufacturer's protocol. As shown in FIG. 5, DD-M. vaccae was found tostimulate significant levels of IL-10 production, suggesting that thismay be the mechanism for the therapeutic action of DD-M. vaccae inpsoriasis. The PBS control did not stimulate THP-1 cells to produceIL-10.

[0235] Although the present invention has been described in some detailby way of illustration and example for purposes of clarity ofunderstanding, changes and modifications can be carried out withoutdeparting from the scope of the invention which is intended to belimited only by the scope of the claims.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 194 <210> SEQ ID NO 1<211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (7)...(7) <400>SEQUENCE: 1 Ala Pro Val Gly Pro Gly Xaa Ala Ala Tyr Val Gln Gln Val ProAsp 1 5 10 15 Gly Pro Gly Ser Val Gln Gly Met Ala 20 25 <210> SEQ ID NO2 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <400>SEQUENCE: 2 Met Xaa Asp Gln Leu Lys Val Asn Asp Asp 1 5 10 <210> SEQ IDNO 3 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2)<400> SEQUENCE: 3 Met Xaa Pro Val Pro Val Ala Thr Ala Ala Tyr 1 5 10<210> SEQ ID NO 4 <211> LENGTH: 21 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 4 Thr Pro Ala Pro Ala Pro Pro ProTyr Val Asp His Val Glu Gln Ala 1 5 10 15 Lys Phe Gly Asp Leu 20 <210>SEQ ID NO 5 <211> LENGTH: 29 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (25)...(25) <400> SEQUENCE: 5 Met Gln Ala Phe Asn Ala Asp AlaTyr Ala Phe Ala Lys Arg Glu Lys 1 5 10 15 Val Ser Leu Ala Pro Gly ValPro Xaa Val Phe Glu Thr 20 25 <210> SEQ ID NO 6 <211> LENGTH: 21 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (6)...(6) <400> SEQUENCE: 6 Met Ala AspPro Asn Xaa Ala Ile Leu Gln Val Ser Lys Thr Thr Arg 1 5 10 15 Gly GlyGln Ala Ala 20 <210> SEQ ID NO 7 <211> LENGTH: 11 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 7 Met Pro Ile Leu Gln ValSer Gln Thr Gly Arg 1 5 10 <210> SEQ ID NO 8 <211> LENGTH: 14 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <221> NAME/KEY: UNSURE <222>LOCATION: (6)...(6) <400> SEQUENCE: 8 Met Xaa Asp Pro Ile Xaa Leu GlnLeu Gln Val Ser Ser Thr 1 5 10 <210> SEQ ID NO 9 <211> LENGTH: 16 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 9 Lys AlaThr Tyr Val Gln Gly Gly Leu Gly Arg Ile Glu Ala Arg Val 1 5 10 15 <210>SEQ ID NO 10 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (2)...(2) <400> SEQUENCE: 10 Lys Xaa Gly Leu Ala Asp Leu AlaPro 1 5 <210> SEQ ID NO 11 <211> LENGTH: 14 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (12)...(12) <223> OTHER INFORMATION: Residue can beeither Glu or Ile <221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <400>SEQUENCE: 11 Lys Xaa Tyr Ala Leu Ala Leu Met Ser Ala Val Xaa Ala Ala 1 510 <210> SEQ ID NO 12 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (10)...(10) <400> SEQUENCE: 12 Lys Asn Pro Gln Val Ser Asp GluLeu Xaa Thr 1 5 10 <210> SEQ ID NO 13 <211> LENGTH: 21 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (9)...(9) <400> SEQUENCE: 13 Ala Pro Ala Pro AlaAla Pro Ala Xaa Gly Asp Pro Ala Ala Val Val 1 5 10 15 Ala Ala Met SerThr 20 <210> SEQ ID NO 14 <211> LENGTH: 15 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (5)...(5) <400> SEQUENCE: 14 Glu Ala Glu Val Xaa Tyr LeuGly Gln Pro Gly Glu Leu Val Asn 1 5 10 15 <210> SEQ ID NO 15 <211>LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220>FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <223> OTHERINFORMATION: Residue can be either Gly or Ala <221> NAME/KEY: UNSURE<222> LOCATION: (15)...(15) <223> OTHER INFORMATION: Residue can beeither Pro or Ala <221> NAME/KEY: UNSURE <222> LOCATION: (7)...(7) <400>SEQUENCE: 15 Ala Xaa Val Val Pro Pro Xaa Gly Pro Pro Ala Pro Gly Ala Xaa1 5 10 15 <210> SEQ ID NO 16 <211> LENGTH: 15 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 16 Ala Pro Ala Pro AspLeu Gln Gly Pro Leu Val Ser Thr Leu Ser 1 5 10 15 <210> SEQ ID NO 17<211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 17 Ala Thr Pro Asp Trp Ser Gly Arg Tyr Thr Val Val ThrPhe Ala Ser 1 5 10 15 Asp Lys Leu Gly Thr Ser Val Ala Ala 20 25 <210>SEQ ID NO 18 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (15)...(15) <223> OTHER INFORMATION: Residue can be either Alaor Arg <221> NAME/KEY: UNSURE <222> LOCATION: (23)...(23) <223> OTHERINFORMATION: Residue can be either Val or Leu <221> NAME/KEY: UNSURE<222> LOCATION: (16)...(16) <400> SEQUENCE: 18 Ala Pro Pro Tyr Asp AspArg Gly Tyr Val Asp Ser Thr Ala Xaa Xaa 1 5 10 15 Ala Ser Pro Pro ThrLeu Xaa Val Val 20 25 <210> SEQ ID NO 19 <211> LENGTH: 8 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 19 Glu Pro Glu GlyVal Ala Pro Pro 1 5 <210> SEQ ID NO 20 <211> LENGTH: 25 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (21)...(22) <400> SEQUENCE: 20 Glu Pro Ala GlyIle Pro Ala Gly Phe Pro Asp Val Ser Ala Tyr Ala 1 5 10 15 Ala Val AspPro Xaa Xaa Tyr Val Val 20 25 <210> SEQ ID NO 21 <211> LENGTH: 15 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (7)...(7) <400> SEQUENCE: 21 Ala ProVal Gly Pro Gly Xaa Ala Ala Tyr Val Gln Gln Val Pro 1 5 10 15 <210> SEQID NO 22 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 22 Phe Ser Arg Pro Gly Leu Pro Val Glu Tyr LeuMet Val Pro Ser 1 5 10 15 <210> SEQ ID NO 23 <211> LENGTH: 19 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 23 PheSer Arg Pro Gly Leu Pro Val Glu Tyr Leu Met Val Pro Ser Pro 1 5 10 15Ser Met Gly <210> SEQ ID NO 24 <211> LENGTH: 15 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 24 Phe Ser Arg Pro GlyLeu Pro Val Glu Tyr Leu Asp Val Phe Ser 1 5 10 15 <210> SEQ ID NO 25<211> LENGTH: 14 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (1)...(2) <400>SEQUENCE: 25 Xaa Xaa Thr Gly Leu His Arg Leu Arg Met Met Val Pro Asn 1 510 <210> SEQ ID NO 26 <211> LENGTH: 20 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (16)...(16) <223> OTHER INFORMATION: Residue can be either Seror Val <221> NAME/KEY: UNSURE <222> LOCATION: (17)...(17) <223> OTHERINFORMATION: Residue can be either Gln or Val <400> SEQUENCE: 26 Val ProAla Asp Pro Val Gly Ala Ala Ala Gln Ala Glu Pro Ala Xaa 1 5 10 15 XaaArg Ile Asp 20 <210> SEQ ID NO 27 <211> LENGTH: 14 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (4)...(4) <223> OTHER INFORMATION: Residue can be eitherTyr or Pro <221> NAME/KEY: UNSURE <222> LOCATION: (8)...(8) <223> OTHERINFORMATION: Residue can be either Val or Gly <221> NAME/KEY: UNSURE<222> LOCATION: (9)...(9) <223> OTHER INFORMATION: Residue can be eitherIle or Tyr <221> NAME/KEY: UNSURE <222> LOCATION: (3)...(3) <400>SEQUENCE: 27 Asp Pro Xaa Xaa Asp Ile Glu Xaa Xaa Phe Ala Arg Gly Thr 1 510 <210> SEQ ID NO 28 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 28 Ala Pro Ser Leu Ser Val Ser AspTyr Ala Arg Asp Ala Gly Phe 1 5 10 15 <210> SEQ ID NO 29 <211> LENGTH:16 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <223> OTHERINFORMATION: Residue can be either Leu or Pro <221> NAME/KEY: UNSURE<222> LOCATION: (1)...(1) <221> NAME/KEY: UNSURE <222> LOCATION:(5)...(5) <221> NAME/KEY: UNSURE <222> LOCATION: (7)...(7) <221>NAME/KEY: UNSURE <222> LOCATION: (10)...(10) <400> SEQUENCE: 29 Xaa XaaLeu Ala Xaa Ala Xaa Leu Gly Xaa Thr Val Asp Ala Asp Gln 1 5 10 15 <210>SEQ ID NO 30 <211> LENGTH: 330 <212> TYPE: PRT <213> ORGANISM:Mycobacterium leprae <400> SEQUENCE: 30 Met Lys Phe Val Asp Arg Phe ArgGly Ala Val Ala Gly Met Leu Arg 1 5 10 15 Arg Leu Val Val Glu Ala MetGly Val Ala Leu Leu Ser Ala Leu Ile 20 25 30 Gly Val Val Gly Ser Ala ProAla Glu Ala Phe Ser Arg Pro Gly Leu 35 40 45 Pro Val Glu Tyr Leu Gln ValPro Ser Pro Ser Met Gly Arg Asp Ile 50 55 60 Lys Val Gln Phe Gln Asn GlyGly Ala Asn Ser Pro Ala Leu Tyr Leu 65 70 75 80 Leu Asp Gly Leu Arg AlaGln Asp Asp Phe Ser Gly Trp Asp Ile Asn 85 90 95 Thr Thr Ala Phe Glu TrpTyr Tyr Gln Ser Gly Ile Ser Val Val Met 100 105 110 Pro Val Gly Gly GlnSer Ser Phe Tyr Ser Asp Trp Tyr Ser Pro Ala 115 120 125 Cys Gly Lys AlaGly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu Thr 130 135 140 Ser Glu LeuPro Glu Tyr Leu Gln Ser Asn Lys Gln Ile Lys Pro Thr 145 150 155 160 GlySer Ala Ala Val Gly Leu Ser Met Ala Gly Leu Ser Ala Leu Thr 165 170 175Leu Ala Ile Tyr His Pro Asp Gln Phe Ile Tyr Val Gly Ser Met Ser 180 185190 Gly Leu Leu Asp Pro Ser Asn Ala Met Gly Pro Ser Leu Ile Gly Leu 195200 205 Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala Ala Asp Met Trp Gly Pro210 215 220 Ser Thr Asp Pro Ala Trp Lys Arg Asn Asp Pro Thr Val Asn ValGly 225 230 235 240 Thr Leu Ile Ala Asn Asn Thr Arg Ile Trp Met Tyr CysGly Asn Gly 245 250 255 Lys Pro Thr Glu Leu Gly Gly Asn Asn Leu Pro AlaLys Leu Leu Glu 260 265 270 Gly Leu Val Arg Thr Ser Asn Ile Lys Phe GlnAsp Gly Tyr Asn Ala 275 280 285 Gly Gly Gly His Asn Ala Val Phe Asn PhePro Asp Ser Gly Thr His 290 295 300 Ser Trp Glu Tyr Trp Gly Glu Gln LeuAsn Asp Met Lys Pro Asp Leu 305 310 315 320 Gln Gln Tyr Leu Gly Ala ThrPro Gly Ala 325 330 <210> SEQ ID NO 31 <211> LENGTH: 327 <212> TYPE: PRT<213> ORGANISM: Mycobacterium leprae <400> SEQUENCE: 31 Met Ile Asp ValSer Gly Lys Ile Arg Ala Trp Gly Arg Trp Leu Leu 1 5 10 15 Val Gly AlaAla Ala Thr Leu Pro Ser Leu Ile Ser Leu Ala Gly Gly 20 25 30 Ala Ala ThrAla Ser Ala Phe Ser Arg Pro Gly Leu Pro Val Glu Tyr 35 40 45 Leu Gln ValPro Ser Glu Ala Met Gly Arg Thr Ile Lys Val Gln Phe 50 55 60 Gln Asn GlyGly Asn Gly Ser Pro Ala Val Tyr Leu Leu Asp Gly Leu 65 70 75 80 Arg AlaGln Asp Asp Tyr Asn Gly Trp Asp Ile Asn Thr Ser Ala Phe 85 90 95 Glu TrpTyr Tyr Gln Ser Gly Leu Ser Val Val Met Pro Val Gly Gly 100 105 110 GlnSer Ser Phe Tyr Ser Asp Trp Tyr Ser Pro Ala Cys Gly Lys Ala 115 120 125Gly Cys Thr Thr Tyr Lys Trp Glu Thr Phe Leu Thr Ser Glu Leu Pro 130 135140 Lys Trp Leu Ser Ala Asn Arg Ser Val Lys Ser Thr Gly Ser Ala Val 145150 155 160 Val Gly Leu Ser Met Ala Gly Ser Ser Ala Leu Ile Leu Ala AlaTyr 165 170 175 His Pro Asp Gln Phe Ile Tyr Ala Gly Ser Leu Ser Ala LeuMet Asp 180 185 190 Ser Ser Gln Gly Ile Glu Pro Gln Leu Ile Gly Leu AlaMet Gly Asp 195 200 205 Ala Gly Gly Tyr Lys Ala Ala Asp Met Trp Gly ProPro Asn Asp Pro 210 215 220 Ala Trp Gln Arg Asn Asp Pro Ile Leu Gln AlaGly Lys Leu Val Ala 225 230 235 240 Asn Asn Thr His Leu Trp Val Tyr CysGly Asn Gly Thr Pro Ser Glu 245 250 255 Leu Gly Gly Thr Asn Val Pro AlaGlu Phe Leu Glu Asn Phe Val His 260 265 270 Gly Ser Asn Leu Lys Phe GlnAsp Ala Tyr Asn Gly Ala Gly Gly His 275 280 285 Asn Ala Val Phe Asn LeuAsn Ala Asp Gly Thr His Ser Trp Glu Tyr 290 295 300 Trp Gly Ala Gln LeuAsn Ala Met Lys Pro Asp Leu Gln Asn Thr Leu 305 310 315 320 Met Ala ValPro Arg Ser Gly 325 <210> SEQ ID NO 32 <211> LENGTH: 338 <212> TYPE: PRT<213> ORGANISM: Mycobacterium tuberculosis <400> SEQUENCE: 32 Met GlnLeu Val Asp Arg Val Arg Gly Ala Val Thr Gly Met Ser Arg 1 5 10 15 ArgLeu Val Val Gly Ala Val Gly Ala Ala Leu Val Ser Gly Leu Val 20 25 30 GlyAla Val Gly Gly Thr Ala Thr Ala Gly Ala Phe Ser Arg Pro Gly 35 40 45 LeuPro Val Glu Tyr Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp 50 55 60 IleLys Val Gln Phe Gln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr 65 70 75 80Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser Gly Trp Asp Ile 85 90 95Asn Thr Pro Ala Phe Glu Trp Tyr Asp Gln Ser Gly Leu Ser Val Val 100 105110 Met Pro Val Gly Gly Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Gln Pro 115120 125 Ala Cys Gly Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu130 135 140 Thr Ser Glu Leu Pro Gly Trp Leu Gln Ala Asn Arg His Val LysPro 145 150 155 160 Thr Gly Ser Ala Val Val Gly Leu Ser Met Ala Ala SerSer Ala Leu 165 170 175 Thr Leu Ala Ile Tyr His Pro Gln Gln Phe Val TyrAla Gly Ala Met 180 185 190 Ser Gly Leu Leu Asp Pro Ser Gln Ala Met GlyPro Thr Leu Ile Gly 195 200 205 Leu Ala Met Gly Asp Ala Gly Gly Tyr LysAla Ser Asp Met Trp Gly 210 215 220 Pro Lys Glu Asp Pro Ala Trp Gln ArgAsn Asp Pro Leu Leu Asn Val 225 230 235 240 Gly Lys Leu Ile Ala Asn AsnThr Arg Val Trp Val Tyr Cys Gly Asn 245 250 255 Gly Lys Pro Ser Asp LeuGly Gly Asn Asn Leu Pro Ala Lys Phe Leu 260 265 270 Glu Gly Phe Val ArgThr Ser Asn Ile Lys Phe Gln Asp Ala Tyr Asn 275 280 285 Ala Gly Gly GlyHis Asn Gly Val Phe Asp Phe Pro Asp Ser Gly Thr 290 295 300 His Ser TrpGlu Tyr Trp Gly Ala Gln Leu Asn Ala Met Lys Pro Asp 305 310 315 320 LeuGln Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly Pro Ala Pro Gln 325 330 335Gly Ala <210> SEQ ID NO 33 <211> LENGTH: 325 <212> TYPE: PRT <213>ORGANISM: Mycobacterium tuberculosis <400> SEQUENCE: 33 Met Thr Asp ValSer Arg Lys Ile Arg Ala Trp Gly Arg Arg Leu Met 1 5 10 15 Ile Gly ThrAla Ala Ala Val Val Leu Pro Gly Leu Val Gly Leu Ala 20 25 30 Gly Gly AlaAla Thr Ala Gly Ala Phe Ser Arg Pro Gly Leu Pro Val 35 40 45 Glu Tyr LeuGln Val Pro Ser Pro Ser Met Gly Arg Asp Ile Lys Val 50 55 60 Gln Phe GlnSer Gly Gly Asn Asn Ser Pro Ala Val Tyr Leu Leu Asp 65 70 75 80 Gly LeuArg Ala Gln Asp Asp Tyr Asn Gly Trp Asp Ile Asn Thr Pro 85 90 95 Ala PheGlu Trp Tyr Tyr Gln Ser Gly Leu Ser Ile Val Met Pro Val 100 105 110 GlyGly Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Ser Pro Ala Cys Gly 115 120 125Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu Thr Ser Glu 130 135140 Leu Pro Gln Trp Leu Ser Ala Asn Arg Ala Val Lys Pro Thr Gly Ser 145150 155 160 Ala Ala Ile Gly Leu Ser Met Ala Gly Ser Ser Ala Met Ile LeuAla 165 170 175 Ala Tyr His Pro Gln Gln Phe Ile Tyr Ala Gly Ser Leu SerAla Leu 180 185 190 Leu Asp Pro Ser Gln Gly Met Gly Pro Ser Leu Ile GlyLeu Ala Met 195 200 205 Gly Asp Ala Gly Gly Tyr Lys Ala Ala Asp Met TrpGly Pro Ser Ser 210 215 220 Asp Pro Ala Trp Glu Arg Asn Asp Pro Thr GlnGln Ile Pro Lys Leu 225 230 235 240 Val Ala Asn Asn Thr Arg Leu Trp ValTyr Cys Gly Asn Gly Thr Pro 245 250 255 Asn Glu Leu Gly Gly Ala Asn IlePro Ala Glu Phe Leu Glu Asn Phe 260 265 270 Val Arg Ser Ser Asn Leu LysPhe Gln Asp Ala Tyr Asn Ala Ala Gly 275 280 285 Gly His Asn Ala Val PheAsn Phe Pro Pro Asn Gly Thr His Ser Trp 290 295 300 Glu Tyr Trp Gly AlaGln Leu Asn Ala Met Lys Gly Asp Leu Gln Ser 305 310 315 320 Ser Leu GlyAla Gly 325 <210> SEQ ID NO 34 <211> LENGTH: 338 <212> TYPE: PRT <213>ORGANISM: Mycobacterium bovis <400> SEQUENCE: 34 Met Gln Leu Val Asp ArgVal Arg Gly Ala Val Thr Gly Met Ser Arg 1 5 10 15 Arg Leu Val Val GlyAla Val Gly Ala Ala Leu Val Ser Gly Leu Val 20 25 30 Gly Ala Val Gly GlyThr Ala Thr Ala Gly Ala Phe Ser Arg Pro Gly 35 40 45 Leu Pro Val Glu TyrLeu Gln Val Pro Ser Pro Ser Met Gly Arg Asp 50 55 60 Ile Lys Val Gln PheGln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr 65 70 75 80 Leu Leu Asp GlyLeu Arg Ala Gln Asp Asp Phe Ser Gly Trp Asp Ile 85 90 95 Asn Thr Pro AlaPhe Glu Trp Tyr Asp Gln Ser Gly Leu Ser Val Val 100 105 110 Met Pro ValGly Gly Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Gln Pro 115 120 125 Ala CysGly Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu 130 135 140 ThrSer Glu Leu Pro Gly Trp Leu Gln Ala Asn Arg His Val Lys Pro 145 150 155160 Thr Gly Ser Ala Val Val Gly Leu Ser Met Ala Ala Ser Ser Ala Leu 165170 175 Thr Leu Ala Ile Tyr His Pro Gln Gln Phe Val Tyr Ala Gly Ala Met180 185 190 Ser Gly Leu Leu Asp Pro Ser Gln Ala Met Gly Pro Thr Leu IleGly 195 200 205 Leu Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala Ser Asp MetTrp Gly 210 215 220 Pro Lys Glu Asp Pro Ala Trp Gln Arg Asn Asp Pro LeuLeu Asn Val 225 230 235 240 Gly Lys Leu Ile Ala Asn Asn Thr Arg Val TrpVal Tyr Cys Gly Asn 245 250 255 Gly Lys Pro Ser Asp Leu Gly Gly Asn AsnLeu Pro Ala Lys Phe Leu 260 265 270 Glu Gly Phe Val Arg Thr Ser Asn IleLys Phe Gln Asp Ala Tyr Asn 275 280 285 Ala Gly Gly Gly His Asn Gly ValPhe Asp Phe Pro Asp Ser Gly Thr 290 295 300 His Ser Trp Glu Tyr Trp GlyAla Gln Leu Asn Ala Met Lys Pro Asp 305 310 315 320 Leu Gln Arg Ala LeuGly Ala Thr Pro Asn Thr Gly Pro Ala Pro Gln 325 330 335 Gly Ala <210>SEQ ID NO 35 <211> LENGTH: 323 <212> TYPE: PRT <213> ORGANISM:Mycobacterium bovis <400> SEQUENCE: 35 Met Thr Asp Val Ser Arg Lys IleArg Ala Trp Gly Arg Arg Leu Met 1 5 10 15 Ile Gly Thr Ala Ala Ala ValVal Leu Pro Gly Leu Val Gly Leu Ala 20 25 30 Gly Gly Ala Ala Thr Ala GlyAla Phe Ser Arg Pro Gly Leu Pro Val 35 40 45 Glu Tyr Leu Gln Val Pro SerPro Ser Met Gly Arg Asp Ile Lys Val 50 55 60 Gln Phe Gln Ser Gly Gly AsnAsn Ser Pro Ala Val Tyr Leu Leu Asp 65 70 75 80 Gly Leu Arg Ala Gln AspAsp Tyr Asn Gly Trp Asp Ile Asn Thr Pro 85 90 95 Ala Phe Glu Trp Tyr TyrGln Ser Gly Leu Ser Ile Val Met Pro Val 100 105 110 Gly Gly Gln Ser SerPhe Tyr Ser Asp Trp Tyr Ser Pro Ala Cys Gly 115 120 125 Lys Ala Gly CysGln Thr Tyr Lys Trp Glu Thr Leu Leu Thr Ser Glu 130 135 140 Leu Pro GlnTrp Leu Ser Ala Asn Arg Ala Val Lys Pro Thr Gly Ser 145 150 155 160 AlaAla Ile Gly Leu Ser Met Ala Gly Ser Ser Ala Met Ile Leu Ala 165 170 175Ala Tyr His Pro Gln Gln Phe Ile Tyr Ala Gly Ser Leu Ser Ala Leu 180 185190 Leu Asp Pro Ser Gln Gly Met Gly Leu Ile Gly Leu Ala Met Gly Asp 195200 205 Ala Gly Gly Tyr Lys Ala Ala Asp Met Trp Gly Pro Ser Ser Asp Pro210 215 220 Ala Trp Glu Arg Asn Asp Pro Thr Gln Gln Ile Pro Lys Leu ValAla 225 230 235 240 Asn Asn Thr Arg Leu Trp Val Tyr Cys Gly Asn Gly ThrPro Asn Glu 245 250 255 Leu Gly Gly Ala Asn Ile Pro Ala Glu Phe Leu GluAsn Phe Val Arg 260 265 270 Ser Ser Asn Leu Lys Phe Gln Asp Ala Tyr LysPro Ala Gly Gly His 275 280 285 Asn Ala Val Phe Asn Phe Pro Pro Asn GlyThr His Ser Trp Glu Tyr 290 295 300 Trp Gly Ala Gln Leu Asn Ala Met LysGly Asp Leu Gln Ser Ser Leu 305 310 315 320 Gly Ala Gly <210> SEQ ID NO36 <211> LENGTH: 333 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumleprae <400> SEQUENCE: 36 Met Lys Phe Leu Gln Gln Met Arg Lys Leu PheGly Leu Ala Ala Lys 1 5 10 15 Phe Pro Ala Arg Leu Thr Ile Ala Val IleGly Thr Ala Leu Leu Ala 20 25 30 Gly Leu Val Gly Val Val Gly Asp Thr AlaIle Ala Val Ala Phe Ser 35 40 45 Lys Pro Gly Leu Pro Val Glu Tyr Leu GlnVal Pro Ser Pro Ser Met 50 55 60 Gly His Asp Ile Lys Ile Gln Phe Gln GlyGly Gly Gln His Ala Val 65 70 75 80 Tyr Leu Leu Asp Gly Leu Arg Ala GlnGlu Asp Tyr Asn Gly Trp Asp 85 90 95 Ile Asn Thr Pro Ala Phe Glu Glu TyrTyr His Ser Gly Leu Ser Val 100 105 110 Ile Met Pro Val Gly Gly Gln SerSer Phe Tyr Ser Asn Trp Tyr Gln 115 120 125 Pro Ser Gln Gly Asn Gly GlnHis Tyr Thr Tyr Lys Trp Glu Thr Phe 130 135 140 Leu Thr Gln Glu Met ProSer Trp Leu Gln Ala Asn Lys Asn Val Leu 145 150 155 160 Pro Thr Gly AsnAla Ala Val Gly Leu Ser Met Ser Gly Ser Ser Ala 165 170 175 Leu Ile LeuAla Ser Tyr Tyr Pro Gln Gln Phe Pro Tyr Ala Ala Ser 180 185 190 Leu SerGly Phe Leu Asn Pro Ser Glu Gly Trp Trp Pro Thr Met Ile 195 200 205 GlyLeu Ala Met Asn Asp Ser Gly Gly Tyr Asn Ala Asn Ser Met Trp 210 215 220Gly Pro Ser Thr Asp Pro Ala Trp Lys Arg Asn Asp Pro Met Val Gln 225 230235 240 Ile Pro Arg Leu Val Ala Asn Asn Thr Arg Ile Trp Val Tyr Cys Gly245 250 255 Asn Gly Ala Pro Asn Glu Leu Gly Gly Asp Asn Ile Pro Ala LysPhe 260 265 270 Leu Glu Ser Leu Thr Leu Ser Thr Asn Glu Ile Phe Gln AsnThr Tyr 275 280 285 Ala Ala Ser Gly Gly Arg Asn Gly Val Phe Asn Phe ProPro Asn Gly 290 295 300 Thr His Ser Trp Pro Tyr Trp Asn Gln Gln Leu ValAla Met Lys Pro 305 310 315 320 Asp Ile Gln Gln Ile Leu Asn Gly Ser AsnAsn Asn Ala 325 330 <210> SEQ ID NO 37 <211> LENGTH: 340 <212> TYPE: PRT<213> ORGANISM: Mycobacterium tuberculosis <400> SEQUENCE: 37 Met ThrPhe Phe Glu Gln Val Arg Arg Leu Arg Ser Ala Ala Thr Thr 1 5 10 15 LeuPro Arg Arg Val Ala Ile Ala Ala Met Gly Ala Val Leu Val Tyr 20 25 30 GlyLeu Val Gly Thr Phe Gly Gly Pro Ala Thr Ala Gly Ala Phe Ser 35 40 45 ArgPro Gly Leu Pro Val Glu Tyr Leu Gln Val Pro Ser Ala Ser Met 50 55 60 GlyArg Asp Ile Lys Val Gln Phe Gln Gly Gly Gly Pro His Ala Val 65 70 75 80Tyr Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Tyr Asn Gly Trp Asp 85 90 95Ile Asn Thr Pro Ala Phe Glu Glu Tyr Tyr Gln Ser Gly Leu Ser Val 100 105110 Ile Met Pro Val Gly Gly Gln Ser Ser Phe Tyr Thr Asp Trp Tyr Gln 115120 125 Pro Ser Gln Ser Asn Gly Gln Asn Tyr Thr Tyr Lys Trp Glu Thr Phe130 135 140 Leu Thr Arg Glu Met Pro Ala Trp Leu Gln Ala Asn Lys Gly ValSer 145 150 155 160 Pro Thr Gly Asn Ala Ala Val Gly Leu Ser Met Ser GlyGly Ser Ala 165 170 175 Leu Ile Leu Ala Ala Tyr Tyr Pro Gln Gln Phe ProTyr Ala Ala Ser 180 185 190 Leu Ser Gly Phe Leu Asn Pro Ser Glu Gly TrpTrp Pro Thr Leu Ile 195 200 205 Gly Leu Ala Met Asn Asp Ser Gly Gly TyrAsn Ala Asn Ser Met Trp 210 215 220 Gly Pro Ser Ser Asp Pro Ala Trp LysArg Asn Asp Pro Met Val Gln 225 230 235 240 Ile Pro Arg Leu Val Ala AsnAsn Thr Arg Ile Trp Val Tyr Cys Gly 245 250 255 Asn Gly Thr Pro Ser AspLeu Gly Gly Asp Asn Ile Pro Ala Lys Phe 260 265 270 Leu Glu Gly Leu ThrLeu Arg Thr Asn Gln Thr Phe Arg Asp Thr Tyr 275 280 285 Ala Ala Asp GlyGly Arg Asn Gly Val Phe Asn Phe Pro Pro Asn Gly 290 295 300 Thr His SerTrp Pro Tyr Trp Asn Glu Gln Leu Val Ala Met Lys Ala 305 310 315 320 AspIle Gln His Val Leu Asn Gly Ala Thr Pro Pro Ala Ala Pro Ala 325 330 335Ala Pro Ala Ala 340 <210> SEQ ID NO 38 <211> LENGTH: 20 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Probe made in a lab <400> SEQUENCE: 38 agcggctgggacatcaacac 20 <210> SEQ ID NO 39 <211> LENGTH: 20 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Probe made in a lab <400> SEQUENCE: 39 cagacgcggg tgttgttggc 20 <210>SEQ ID NO 40 <211> LENGTH: 1211 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 40 ggtaccggaa gctggaggat tgacggtatgagacttcttg acaggattcg tgggccttgg 60 gcacgccgtt tcggcgtcgt ggctgtcgcgacagcgatga tgcctgcttt ggtgggcctg 120 gctggagggt cggcgaccgc cggagcattctcccggccag gtctgccggt ggagtacctg 180 atggtgcctt cgccgtcgat ggggcgcgacatcaagatcc agttccagag cggtggcgag 240 aactcgccgg ctctctacct gctcgacggcctgcgtgcgc aggaggactt caacggctgg 300 gacatcaaca ctcaggcttt cgagtggttcctcgacagcg gcatctccgt ggtgatgccg 360 gtcggtggcc agtccagctt ctacaccgactggtacgccc ccgcccgtaa caagggcccg 420 accgtgacct acaagtggga gaccttcctgacccaggagc tcccgggctg gctgcaggcc 480 aaccgcgcgg tcaagccgac cggcagcggccctgtcggtc tgtcgatggc gggttcggcc 540 gcgctgaacc tggcgacctg gcacccggagcagttcatct acgcgggctc gatgtccggc 600 ttcctgaacc cctccgaggg ctggtggccgttcctgatca acatctcgat gggtgacgcc 660 ggcggcttca aggccgacga catgtggggcaagaccgagg ggatcccaac agcggttgga 720 cagcgcaacg atccgatgct gaacatcccgaccctggtcg ccaacaacac ccgtatctgg 780 gtctactgcg gtaacggcca gcccaccgagctcggcggcg gcgacctgcc cgccacgttc 840 ctcgaaggtc tgaccatccg caccaacgagaccttccgcg acaactacat cgccgcgggt 900 ggccacaacg gtgtgttcaa cttcccggccaacggcacgc acaactgggc gtactggggt 960 cgcgagctgc aggcgatgaa gcctgacctgcaggcgcacc ttctctgacg gttgcacgaa 1020 acgaagcccc cggccgattg cggccgagggtttcgtcgtc cggggctact gtggccgaca 1080 taaccgaaat caacgcgatg gtggctcatcaggaacgccg agggggtcat tgcgctacga 1140 cacgaggtgg gcgagcaatc cttcctgcccgacggagagg tcaacatcca cgtcgagtac 1200 tccagcgtga a 1211 <210> SEQ ID NO41 <211> LENGTH: 485 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 41 agcggctggg acatcaacac cgccgccttc gagtggtacgtcgactcggg tctcgcggtg 60 atcatgcccg tcggcgggca gtccagcttc tacagcgactggtacagccc ggcctgcggt 120 aaggccggct gccagaccta caagtgggag acgttcctgacccaggagct gccggcctac 180 ctcgccgcca acaagggggt cgacccgaac cgcaacgcggccgtcggtct gtccatggcc 240 ggttcggcgg cgctgacgct ggcgatctac cacccgcagcagttccagta cgccgggtcg 300 ctgtcgggct acctgaaccc gtccgagggg tggtggccgatgctgatcaa catctcgatg 360 ggtgacgcgg gcggctacaa ggccaacgac atgtggggtccaccgaagga cccgagcagc 420 gcctggaagc gcaacgaccc gatggtcaac atcggcaagctggtggccaa caacaccccc 480 ctctc 485 <210> SEQ ID NO 42 <211> LENGTH:1052 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 42 gttgatgaga aaggtgggtt gtttgccgtt atgaagttca cagagaagtggcggggctcc 60 gcaaaggcgg cgatgcaccg ggtgggcgtt gccgatatgg ccgccgttgcgctgcccgga 120 ctgatcggct tcgccggggg ttcggcaacg gccggggcat tctcccggcccggtcttcct 180 gtcgagtacc tcgacgtgtt ctcgccgtcg atgggccgcg acatccgggtccagttccag 240 ggtggcggta ctcatgcggt ctacctgctc gacggtctgc gtgcccaggacgactacaac 300 ggctgggaca tcaacacccc tgcgttcgag tggttctacg agtccggcttgtcgacgatc 360 atgccggtcg gcggacagtc cagcttctac agcgactggt accagccgtctcggggcaac 420 gggcagaact acacctacaa gtgggagacg ttcctgaccc aggagctgccgacgtggctg 480 gaggccaacc gcggagtgtc gcgcaccggc aacgcgttcg tcggcctgtcgatggcgggc 540 agcgcggcgc tgacctacgc gatccatcac ccgcagcagt tcatctacgcctcgtcgctg 600 tcaggcttcc tgaacccgtc cgagggctgg tggccgatgc tgatcgggctggcgatgaac 660 gacgcaggcg gcttcaacgc cgagagcatg tggggcccgt cctcggacccggcgtggaag 720 cgcaacgacc cgatggtcaa catcaaccag ctggtggcca acaacacccggatctggatc 780 tactgcggca ccggcacccc gtcggagctg gacaccggga ccccgggccagaacctgatg 840 gccgcgcagt tcctcgaagg attcacgttg cggaccaaca tcgccttccgtgacaactac 900 atcgcagccg gcggcaccaa cggtgtcttc aacttcccgg cctcgggcacccacagctgg 960 gggtactggg ggcagcagct gcagcagatg aagcccgaca tccagcgggttctgggagct 1020 caggccaccg cctagccacc caccccacac cc 1052 <210> SEQ ID NO43 <211> LENGTH: 326 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 43 Met Arg Leu Leu Asp Arg Ile Arg Gly Pro TrpAla Arg Arg Phe Gly 1 5 10 15 Val Val Ala Val Ala Thr Ala Met Met ProAla Leu Val Gly Leu Ala 20 25 30 Gly Gly Ser Ala Thr Ala Gly Ala Phe SerArg Pro Gly Leu Pro Val 35 40 45 Glu Tyr Leu Met Val Pro Ser Pro Ser MetGly Arg Asp Ile Lys Ile 50 55 60 Gln Phe Gln Ser Gly Gly Glu Asn Ser ProAla Leu Tyr Leu Leu Asp 65 70 75 80 Gly Leu Arg Ala Gln Glu Asp Phe AsnGly Trp Asp Ile Asn Thr Gln 85 90 95 Ala Phe Glu Trp Phe Leu Asp Ser GlyIle Ser Val Val Met Pro Val 100 105 110 Gly Gly Gln Ser Ser Phe Tyr ThrAsp Trp Tyr Ala Pro Ala Arg Asn 115 120 125 Lys Gly Pro Thr Val Thr TyrLys Trp Glu Thr Phe Leu Thr Gln Glu 130 135 140 Leu Pro Gly Trp Leu GlnAla Asn Arg Ala Val Lys Pro Thr Gly Ser 145 150 155 160 Gly Pro Val GlyLeu Ser Met Ala Gly Ser Ala Ala Leu Asn Leu Ala 165 170 175 Thr Trp HisPro Glu Gln Phe Ile Tyr Ala Gly Ser Met Ser Gly Phe 180 185 190 Leu AsnPro Ser Glu Gly Trp Trp Pro Phe Leu Ile Asn Ile Ser Met 195 200 205 GlyAsp Ala Gly Gly Phe Lys Ala Asp Asp Met Trp Gly Lys Thr Glu 210 215 220Gly Ile Pro Thr Ala Val Gly Gln Arg Asn Asp Pro Met Leu Asn Ile 225 230235 240 Pro Thr Leu Val Ala Asn Asn Thr Arg Ile Trp Val Tyr Cys Gly Asn245 250 255 Gly Gln Pro Thr Glu Leu Gly Gly Gly Asp Leu Pro Ala Thr PheLeu 260 265 270 Glu Gly Leu Thr Ile Arg Thr Asn Glu Thr Phe Arg Asp AsnTyr Ile 275 280 285 Ala Ala Gly Gly His Asn Gly Val Phe Asn Phe Pro AlaAsn Gly Thr 290 295 300 His Asn Trp Ala Tyr Trp Gly Arg Glu Leu Gln AlaMet Lys Pro Asp 305 310 315 320 Leu Gln Ala His Leu Leu 325 <210> SEQ IDNO 44 <211> LENGTH: 161 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 44 Ser Gly Trp Asp Ile Asn Thr Ala Ala Phe GluTrp Tyr Val Asp Ser 1 5 10 15 Gly Leu Ala Val Ile Met Pro Val Gly GlyGln Ser Ser Phe Tyr Ser 20 25 30 Asp Trp Tyr Ser Pro Ala Cys Gly Lys AlaGly Cys Gln Thr Tyr Lys 35 40 45 Trp Glu Thr Phe Leu Thr Gln Glu Leu ProAla Tyr Leu Ala Ala Asn 50 55 60 Lys Gly Val Asp Pro Asn Arg Asn Ala AlaVal Gly Leu Ser Met Ala 65 70 75 80 Gly Ser Ala Ala Leu Thr Leu Ala IleTyr His Pro Gln Gln Phe Gln 85 90 95 Tyr Ala Gly Ser Leu Ser Gly Tyr LeuAsn Pro Ser Glu Gly Trp Trp 100 105 110 Pro Met Leu Ile Asn Ile Ser MetGly Asp Ala Gly Gly Tyr Lys Ala 115 120 125 Asn Asp Met Trp Gly Pro ProLys Asp Pro Ser Ser Ala Trp Lys Arg 130 135 140 Asn Asp Pro Met Val AsnIle Gly Lys Leu Val Ala Asn Asn Thr Pro 145 150 155 160 Leu <210> SEQ IDNO 45 <211> LENGTH: 334 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 45 Met Lys Phe Thr Glu Lys Trp Arg Gly Ser AlaLys Ala Ala Met His 1 5 10 15 Arg Val Gly Val Ala Asp Met Ala Ala ValAla Leu Pro Gly Leu Ile 20 25 30 Gly Phe Ala Gly Gly Ser Ala Thr Ala GlyAla Phe Ser Arg Pro Gly 35 40 45 Leu Pro Val Glu Tyr Leu Asp Val Phe SerPro Ser Met Gly Arg Asp 50 55 60 Ile Arg Val Gln Phe Gln Gly Gly Gly ThrHis Ala Val Tyr Leu Leu 65 70 75 80 Asp Gly Leu Arg Ala Gln Asp Asp TyrAsn Gly Trp Asp Ile Asn Thr 85 90 95 Pro Ala Phe Glu Trp Phe Tyr Glu SerGly Leu Ser Thr Ile Met Pro 100 105 110 Val Gly Gly Gln Ser Ser Phe TyrSer Asp Trp Tyr Gln Pro Ser Arg 115 120 125 Gly Asn Gly Gln Asn Tyr ThrTyr Lys Trp Glu Thr Phe Leu Thr Gln 130 135 140 Glu Leu Pro Thr Trp LeuGlu Ala Asn Arg Gly Val Ser Arg Thr Gly 145 150 155 160 Asn Ala Phe ValGly Leu Ser Met Ala Gly Ser Ala Ala Leu Thr Tyr 165 170 175 Ala Ile HisHis Pro Gln Gln Phe Ile Tyr Ala Ser Ser Leu Ser Gly 180 185 190 Phe LeuAsn Pro Ser Glu Gly Trp Trp Pro Met Leu Ile Gly Leu Ala 195 200 205 MetAsn Asp Ala Gly Gly Phe Asn Ala Glu Ser Met Trp Gly Pro Ser 210 215 220Ser Asp Pro Ala Trp Lys Arg Asn Asp Pro Met Val Asn Ile Asn Gln 225 230235 240 Leu Val Ala Asn Asn Thr Arg Ile Trp Ile Tyr Cys Gly Thr Gly Thr245 250 255 Pro Ser Glu Leu Asp Thr Gly Thr Pro Gly Gln Asn Leu Met AlaAla 260 265 270 Gln Phe Leu Glu Gly Phe Thr Leu Arg Thr Asn Ile Ala PheArg Asp 275 280 285 Asn Tyr Ile Ala Ala Gly Gly Thr Asn Gly Val Phe AsnPhe Pro Ala 290 295 300 Ser Gly Thr His Ser Trp Gly Tyr Trp Gly Gln GlnLeu Gln Gln Met 305 310 315 320 Lys Pro Asp Ile Gln Arg Val Leu Gly AlaGln Ala Thr Ala 325 330 <210> SEQ ID NO 46 <211> LENGTH: 795 <212> TYPE:DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 46 ctgccgcgggtttgccatct cttgggtcct gggtcgggag gccatgttct gggtaacgat 60 ccggtaccgtccggcgatgt gaccaacatg cgaacagcga caacgaagct aggagcggcg 120 ctcggcgcagcagcattggt ggccgccacg gggatggtca gcgcggcgac ggcgaacgcc 180 caggaagggcaccaggtccg ttacacgctc acctcggccg gcgcttacga gttcgacctg 240 ttctatctgacgacgcagcc gccgagcatg caggcgttca acgccgacgc gtatgcgttc 300 gccaagcgggagaaggtcag cctcgccccg ggtgtgccgt gggtcttcga aaccacgatg 360 gccgacccgaactgggcgat ccttcaggtc agcagcacca cccgcggtgg gcaggccgcc 420 ccgaacgcgcactgcgacat cgccgtcgat ggccaggagg tgctcagcca gcacgacgac 480 ccctacaacgtgcggtgcca gctcggtcag tggtgagtca cctcgccgag agtccggcca 540 gcgccggcggcagcggctcg cggtgcagca ccccgaggcg ctgggtcgcg cgggtcagcg 600 cgacgtaaagatcgctggcc ccgcgcggcc cctcggcgag gatctgctcc gggtagacca 660 ccagcacggcgtctaactcc agacccttgg tctgcgtggg tgccaccgcg cccgggacac 720 cgggcgggccgatcaccacg ctggtgccct cccggtccgc ctccgcacgc acgaaatcgt 780 cgatggcaccggcga 795 <210> SEQ ID NO 47 <211> LENGTH: 142 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 47 Met Arg Thr Ala ThrThr Lys Leu Gly Ala Ala Leu Gly Ala Ala Ala 1 5 10 15 Leu Val Ala AlaThr Gly Met Val Ser Ala Ala Thr Ala Asn Ala Gln 20 25 30 Glu Gly His GlnVal Arg Tyr Thr Leu Thr Ser Ala Gly Ala Tyr Glu 35 40 45 Phe Asp Leu PheTyr Leu Thr Thr Gln Pro Pro Ser Met Gln Ala Phe 50 55 60 Asn Ala Asp AlaTyr Ala Phe Ala Lys Arg Glu Lys Val Ser Leu Ala 65 70 75 80 Pro Gly ValPro Trp Val Phe Glu Thr Thr Met Ala Asp Pro Asn Trp 85 90 95 Ala Ile LeuGln Val Ser Ser Thr Thr Arg Gly Gly Gln Ala Ala Pro 100 105 110 Asn AlaHis Cys Asp Ile Ala Val Asp Gly Gln Glu Val Leu Ser Gln 115 120 125 HisAsp Asp Pro Tyr Asn Val Arg Cys Gln Leu Gly Gln Trp 130 135 140 <210>SEQ ID NO 48 <211> LENGTH: 300 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 48 gccagtgcgc caacggtttt catcgatgccgcacacaacc ccggtgggcc ctgcgcttgc 60 cgaaggctgc gcgacgagtt cgacttccggtatctcgtcg gcgtcgtctc ggtgatgggg 120 gacaaggacg tggacgggat ccgccaggacccgggcgtgc cggacgggcg cggtctcgca 180 ctgttcgtct cgggcgacaa ccttcgaaagggtgcggcgc tcaacacgat ccagatcgcc 240 gagctgctgg ccgcccagtt gtaagtgttccgccgaaatt gcattccacg ccgataatcg 300 <210> SEQ ID NO 49 <211> LENGTH:563 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE:49 ggatcctcgg ccggctcaag agtccgcgcc gaggtggatg tgacgctgga cggctacgag 60ttcagtcggg cctgcgaggc gctgtaccac ttcgcctggg acgagttctg cgactggtat 120gtcgagcttg ccaaagtgca actgggtgaa ggtttctcgc acaccacggc cgtgttggcc 180accgtgctcg atgtgctgct caagcttctg cacccggtca tgccgttcgt caccgaggtg 240ctgtggaagg ccctgaccgg gcgggccggc gcgagcgaac gtctgggaaa tgtggagtca 300ctggtcgtcg cggactggcc cacgcccacc ggatacgcgc tggatcaggc tgccgcacaa 360cggatcgccg acacccagaa gttgatcacc gaggtgcgcc ggttccgcag cgatcagggt 420ctggccgacc gccagcgggt gcctgcccgg ttgtccggca tcgacaccgc gggtctggac 480gcccatgtcc cggcggtgcg cgcgctggcc tggcttgacc gagggtgatg agggcttcac 540cgcgtccgaa tcggtcgagg tgc 563 <210> SEQ ID NO 50 <211> LENGTH: 434 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 50gggccgggcc cgaggatgag caagttcgaa gtcgtcaccg ggatggcgtt cgcggctttc 60gccgacgcgc ccatcgacgt cgccgtcgtc gaggtcgggc tcggtggtcg ctgggacgcg 120acgaacgtgg tgaacgcacc ggtcgcggtc atcaccccga tcggggtgga ccacaccgac 180tacctcggtg acacgatcgc cgagatcgcc ggggagaagg ccggaaatca tcacccgcca 240gccgacgacc tggtgccgac cgacaccgtc gccgtgctgg cgcggcaggt tcccgaggcc 300atggaggtgc tgctggccca ggcggtgcgc tcggatgcgg ctgtagcgcg cgaggattcg 360gagtgcgcgg tgctgggccg tcaggtcgcc atcggcggca gctgctccgg ttgcaggggc 420tcggtggcgt ctac 434 <210> SEQ ID NO 51 <211> LENGTH: 438 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 51 ggatcccactcccgcgccgg cggcggccag ctggtacggc cattccagcg tgctgatcga 60 ggtcgacggctaccgcgtgc tggccgaccc ggtgtggagc aacagatgtt cgccctcacg 120 ggcggtcggaccgcagcgca tgcacgacgt cccggtgccg ctggaggcgc ttcccgccgt 180 ggacgcggtggtgatcgcca acgaccacta cgaccacctc gacatcgaca ccatcgtcgc 240 gttggcgcacacccagcggg ccccgttcgt ggtgccgttg ggcatcggcg cacacctgcg 300 caagtggggcgtccccgagg cgcggatcgt cgagttggac tggcacgaag cccaccgcat 360 cgacgacctgacgctggtct gcacccccgc ccggcacttc tccggccggt tgttctcccg 420 cgactcgacgctgtgggc 438 <210> SEQ ID NO 52 <211> LENGTH: 87 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 52 Ala Ser Ala Pro ThrVal Phe Ile Asp Ala Ala His Asn Pro Gly Gly 1 5 10 15 Pro Cys Ala CysArg Arg Leu Arg Asp Glu Phe Asp Phe Arg Tyr Leu 20 25 30 Val Gly Val ValSer Val Met Gly Asp Lys Asp Val Asp Gly Ile Arg 35 40 45 Gln Asp Pro GlyVal Pro Asp Gly Arg Gly Leu Ala Leu Phe Val Ser 50 55 60 Gly Asp Asn LeuArg Lys Gly Ala Ala Leu Asn Thr Ile Gln Ile Ala 65 70 75 80 Glu Leu LeuAla Ala Gln Leu 85 <210> SEQ ID NO 53 <211> LENGTH: 175 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 53 Gly Ser Ser AlaGly Ser Arg Val Arg Ala Glu Val Asp Val Thr Leu 1 5 10 15 Asp Gly TyrGlu Phe Ser Arg Ala Cys Glu Ala Leu Tyr His Phe Ala 20 25 30 Trp Asp GluPhe Cys Asp Trp Tyr Val Glu Leu Ala Lys Val Gln Leu 35 40 45 Gly Glu GlyPhe Ser His Thr Thr Ala Val Leu Ala Thr Val Leu Asp 50 55 60 Val Leu LeuLys Leu Leu His Pro Val Met Pro Phe Val Thr Glu Val 65 70 75 80 Leu TrpLys Ala Leu Thr Gly Arg Ala Gly Ala Ser Glu Arg Leu Gly 85 90 95 Asn ValGlu Ser Leu Val Val Ala Asp Trp Pro Thr Pro Thr Gly Tyr 100 105 110 AlaLeu Asp Gln Ala Ala Ala Gln Arg Ile Ala Asp Thr Gln Lys Leu 115 120 125Ile Thr Glu Val Arg Arg Phe Arg Ser Asp Gln Gly Leu Ala Asp Arg 130 135140 Gln Arg Val Pro Ala Arg Leu Ser Gly Ile Asp Thr Ala Gly Leu Asp 145150 155 160 Ala His Val Pro Ala Val Arg Ala Leu Ala Trp Leu Asp Arg Gly165 170 175 <210> SEQ ID NO 54 <211> LENGTH: 144 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 54 Gly Pro Gly Pro ArgAsn Ser Lys Phe Glu Val Val Thr Gly Met Ala 1 5 10 15 Phe Ala Ala PheAla Asp Ala Pro Ile Asp Val Ala Val Val Glu Val 20 25 30 Gly Leu Gly GlyArg Trp Asp Ala Thr Asn Val Val Asn Ala Pro Val 35 40 45 Ala Val Ile ThrPro Ile Gly Val Asp His Thr Asp Tyr Leu Gly Asp 50 55 60 Thr Ile Ala GluIle Ala Gly Glu Lys Ala Gly Asn His His Pro Pro 65 70 75 80 Ala Asp AspLeu Val Pro Thr Asp Thr Val Ala Val Leu Ala Arg Gln 85 90 95 Val Pro GluAla Asn Glu Val Leu Leu Ala Gln Ala Val Arg Ser Asp 100 105 110 Ala AlaVal Ala Arg Glu Asp Ser Glu Cys Ala Val Leu Gly Arg Gln 115 120 125 ValAla Ile Gly Gly Ser Cys Ser Gly Cys Arg Gly Ser Val Ala Ser 130 135 140<210> SEQ ID NO 55 <211> LENGTH: 145 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 55 Asp Pro Thr Pro Ala Pro Ala AlaAla Ser Trp Tyr Gly His Ser Ser 1 5 10 15 Val Leu Ile Glu Val Asp GlyTyr Arg Val Leu Ala Asp Pro Val Trp 20 25 30 Ser Asn Arg Cys Ser Pro SerArg Ala Val Gly Pro Gln Arg Met His 35 40 45 Asp Val Pro Val Pro Leu GluAla Leu Pro Ala Val Asp Ala Val Val 50 55 60 Ile Ser Asn Asp His Tyr AspHis Leu Asp Ile Asp Thr Ile Val Ala 65 70 75 80 Leu Ala His Thr Gln ArgAla Pro Phe Val Val Pro Leu Gly Ile Gly 85 90 95 Ala His Leu Arg Lys TrpGly Val Pro Glu Ala Arg Ile Val Glu Leu 100 105 110 Asp Trp His Glu AlaHis Arg Ile Asp Asp Leu Thr Leu Val Cys Thr 115 120 125 Pro Ala Arg HisPhe Ser Gly Arg Leu Phe Ser Arg Asp Ser Thr Leu 130 135 140 Trp 145<210> SEQ ID NO 56 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (1)...(1) <223> OTHER INFORMATION: Residue can be either Gly,Ile, Leu or Val <221> NAME/KEY: UNSURE <222> LOCATION: (2)...(2) <223>OTHER INFORMATION: Residue can be either Ile, Leu, Gly, or Ala <221>NAME/KEY: UNSURE <222> LOCATION: (5)...(5) <221> NAME/KEY: UNSURE <222>LOCATION: (9)...(9) <400> SEQUENCE: 56 Xaa Xaa Ala Pro Xaa Gly Asp AlaXaa Arg 1 5 10 <210> SEQ ID NO 57 <211> LENGTH: 8 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (7)...(7) <223> OTHER INFORMATION: Residue can be eitherIle or Leu <400> SEQUENCE: 57 Pro Glu Ala Glu Ala Asn Xaa Arg 1 5 <210>SEQ ID NO 58 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (4)...(4) <223> OTHER INFORMATION: Residue can be either Glnor Gly <221> NAME/KEY: UNSURE <222> LOCATION: (5)...(5) <223> OTHERINFORMATION: Residue can be either Gly or Gln <400> SEQUENCE: 58 Thr AlaAsn Xaa Xaa Glu Tyr Tyr Asp Asn Arg 1 5 10 <210> SEQ ID NO 59 <211>LENGTH: 34 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 59 Asn Ser Pro Arg Ala Glu Ala Glu Ala Asn Leu Arg Gly Tyr PheThr 1 5 10 15 Ala Asn Pro Ala Glu Tyr Tyr Asp Leu Arg Gly Ile Leu AlaPro Ile 20 25 30 Gly Asp <210> SEQ ID NO 60 <211> LENGTH: 20 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 60 ccggtgggcc cgggctgcgc 20<210> SEQ ID NO 61 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 61 tggccggcca ccacgtggta 20 <210> SEQ ID NO 62 <211>LENGTH: 313 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 62 gccggtgggc ccgggctgcg cggaatacgc ggcagccaat cccactgggccggcctcggt 60 gcagggaatg tcgcaggacc cggtcgcggt ggcggcctcg aacaatccggagttgacaac 120 gctgtacggc tgcactgtcg ggccagctca atccgcaagt aaacctggtggacaccctca 180 acagcggtca gtacacggtg ttcgcaccga ccaacgcggc atttagcaagctgccggcat 240 ccacgatcga cgagctcaag accaattcgt cactgctgac cagcatcctgacctaccacg 300 tggtggccgg cca 313 <210> SEQ ID NO 63 <211> LENGTH: 18<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (7)...(17) <400> SEQUENCE: 63 GluPro Ala Gly Pro Leu Pro Xaa Tyr Asn Glu Arg Leu His Thr Leu 1 5 10 15Xaa Gln <210> SEQ ID NO 64 <211> LENGTH: 25 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (21)...(21) <400> SEQUENCE: 64 Gly Leu Asp Asn Glu LeuSer Leu Val Asp Gly Gln Gly Arg Thr Leu 1 5 10 15 Thr Val Gln Gln XaaAsp Thr Phe Leu 20 25 <210> SEQ ID NO 65 <211> LENGTH: 26 <212> TYPE:PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (3)...(3) <221> NAME/KEY: UNSURE <222> LOCATION:(21)...(22) <221> NAME/KEY: UNSURE <222> LOCATION: (24)...(24) <400>SEQUENCE: 65 Asp Pro Xaa Pro Asp Ile Glu Val Glu Phe Ala Arg Gly Thr GlyAla 1 5 10 15 Glu Pro Gly Leu Xaa Xaa Val Xaa Asp Ala 20 25 <210> SEQ IDNO 66 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Made in a lab <400>SEQUENCE: 66 accgccctcg agttctcccg gccaggtctg cc 32 <210> SEQ ID NO 67<211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Made in a lab <400> SEQUENCE: 67aagcacgagc tcagtctctt ccacgcggac gt 32 <210> SEQ ID NO 68 <211> LENGTH:30 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:<223> OTHER INFORMATION: Made in a lab <400> SEQUENCE: 68 catggatccattctcccggc ccggtcttcc 30 <210> SEQ ID NO 69 <211> LENGTH: 26 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 69 tttgaattct aggcggtggcctgagc 26 <210> SEQ ID NO 70 <211> LENGTH: 161 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 70 Ser Gly Trp Asp IleAsn Thr Ala Ala Phe Glu Trp Tyr Val Asp Ser 1 5 10 15 Gly Leu Ala ValIle Met Pro Val Gly Gly Gln Ser Ser Phe Tyr Ser 20 25 30 Asp Trp Tyr SerPro Ala Cys Gly Lys Ala Gly Cys Gln Thr Tyr Lys 35 40 45 Trp Glu Thr PheLeu Thr Gln Glu Leu Pro Ala Tyr Leu Ala Ala Asn 50 55 60 Lys Gly Val AspPro Asn Arg Asn Ala Ala Val Gly Leu Ser Met Ala 65 70 75 80 Gly Ser AlaAla Leu Thr Leu Ala Ile Tyr His Pro Gln Gln Phe Gln 85 90 95 Tyr Ala GlySer Leu Ser Gly Tyr Leu Asn Pro Ser Glu Gly Trp Trp 100 105 110 Pro MetLeu Ile Asn Ile Ser Met Gly Asp Ala Gly Gly Tyr Lys Ala 115 120 125 AsnAsp Met Trp Gly Arg Thr Glu Asp Pro Ser Ser Ala Trp Lys Arg 130 135 140Asn Asp Pro Met Val Asn Ile Gly Lys Leu Val Ala Asn Asn Thr Pro 145 150155 160 Leu <210> SEQ ID NO 71 <211> LENGTH: 33 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Made in a lab <400> SEQUENCE: 71 gagagactcg agaacgccca ggaagggcac cag 33<210> SEQ ID NO 72 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 72 gagagactcg agtgactcac cactgaccga gc 32 <210> SEQID NO 73 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Made in a lab <221>NAME/KEY: unsure <222> LOCATION: (3)...(3) <221> NAME/KEY: unsure <222>LOCATION: (6)...(6) <221> NAME/KEY: unsure <222> LOCATION: (9)...(9)<221> NAME/KEY: unsure <222> LOCATION: (15)...(15) <400> SEQUENCE: 73ggngcngcnc argcngarcc 20 <210> SEQ ID NO 74 <211> LENGTH: 825 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 74ttggatccca ctcccgcgcc ggcggcggcc agctggtacg gccattccag cgtgctgatc 60gaggtcgacg gctaccgcgt gctggccgac ccggtgtgga gcaacagatg ttcgccctca 120cgggcggtcg gaccgcagcg catgcacgac gtcccggtgc cgctggaggc gcttcccgcc 180gtggacgcgg tggtgatcag ccacgaccac tacgaccacc tcgacatcga caccatcgtc 240gcgttggcgc acacccagcg ggccccgttc gtggtgccgt tgggcatcgg cgcacacctg 300cgcaagtggg gcgtccccga ggcgcggatc gtcgagttgg actggcacga agcccaccgc 360atagacgacc tgacgctggt ctgcaccccc gcccggcact tctccggacg gttgttctcc 420cgcgactcga cgctgtgggc gtcgtgggtg gtcaccggct cgtcgcacaa ggcgttcttc 480ggtggcgaca ccggatacac gaagagcttc gccgagatcg gcgacgagta cggtccgttc 540gatctgaccc tgctgccgat cggggcctac catcccgcgt tcgccgacat ccacatgaac 600cccgaggagg cggtgcgcgc ccatctggac ctgaccgagg tggacaacag cctgatggtg 660cccatccact gggcgacatt ccgcctcgcc ccgcatccgt ggtccgagcc cgccgaacgc 720ctgctgaccg ctgccgacgc cgagcgggta cgcctgaccg tgccgattcc cggtcagcgg 780gtggacccgg agtcgacgtt cgacccgtgg tggcggttct gaacc 825 <210> SEQ ID NO 75<211> LENGTH: 273 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 75 Leu Asp Pro Thr Pro Ala Pro Ala Ala Ala Ser Trp TyrGly His Ser 1 5 10 15 Ser Val Leu Ile Glu Val Asp Gly Tyr Arg Val LeuAla Asp Pro Val 20 25 30 Trp Ser Asn Arg Cys Ser Pro Ser Arg Ala Val GlyPro Gln Arg Met 35 40 45 His Asp Val Pro Val Pro Leu Glu Ala Leu Pro AlaVal Asp Ala Val 50 55 60 Val Ile Ser His Asp His Tyr Asp His Leu Asp IleAsp Thr Ile Val 65 70 75 80 Ala Leu Ala His Thr Gln Arg Ala Pro Phe ValVal Pro Leu Gly Ile 85 90 95 Gly Ala His Leu Arg Lys Trp Gly Val Pro GluAla Arg Ile Val Glu 100 105 110 Leu Asp Trp His Glu Ala His Arg Ile AspAsp Leu Thr Leu Val Cys 115 120 125 Thr Pro Ala Arg His Phe Ser Gly ArgLeu Phe Ser Arg Asp Ser Thr 130 135 140 Leu Trp Ala Ser Trp Val Val ThrGly Ser Ser His Lys Ala Phe Phe 145 150 155 160 Gly Gly Asp Thr Gly TyrThr Lys Ser Phe Ala Glu Ile Gly Asp Glu 165 170 175 Tyr Gly Pro Phe AspLeu Thr Leu Leu Pro Ile Gly Ala Tyr His Pro 180 185 190 Ala Phe Ala AspIle His Met Asn Pro Glu Glu Ala Val Arg Ala His 195 200 205 Leu Asp LeuThr Glu Val Asp Asn Ser Leu Met Val Pro Ile His Trp 210 215 220 Ala ThrPhe Arg Leu Ala Pro His Pro Trp Ser Glu Pro Ala Glu Arg 225 230 235 240Leu Leu Thr Ala Ala Asp Ala Glu Arg Val Arg Leu Thr Val Pro Ile 245 250255 Pro Gly Gln Arg Val Asp Pro Glu Ser Thr Phe Asp Pro Trp Trp Arg 260265 270 Phe <210> SEQ ID NO 76 <211> LENGTH: 10 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 76 Ala Lys Thr Ile AlaTyr Asp Glu Glu Ala 1 5 10 <210> SEQ ID NO 77 <211> LENGTH: 337 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 77gatccctaca tcctgctggt cagctccaag gtgtcgaccg tcaaggatct gctcccgctg 60ctggagaagg tcatccaggc cggcaagccg ctgctgatca tcgccgagga cgtcgagggc 120gaggccctgt ccacgctggt ggtcaacaag atccgcggca ccttcaagtc cgtcgccgtc 180aaggctccgg gcttcggtga ccgccgcaag gcgatgctgc aggacatggc catcctcacc 240ggtggtcagg tcgtcagcga aagagtcggg ctgtccctgg agaccgccga cgtctcgctg 300ctgggccagg cccgcaaggt cgtcgtcacc aaggaca 337 <210> SEQ ID NO 78 <211>LENGTH: 112 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 78 Asp Pro Tyr Ile Leu Leu Val Ser Ser Lys Val Ser Thr Val LysAsp 1 5 10 15 Leu Leu Pro Leu Leu Glu Lys Val Ile Gln Ala Gly Lys ProLeu Leu 20 25 30 Ile Ile Ala Glu Asp Val Glu Gly Glu Ala Leu Ser Thr LeuVal Val 35 40 45 Asn Lys Ile Arg Gly Thr Phe Lys Ser Val Ala Val Lys AlaPro Gly 50 55 60 Phe Gly Asp Arg Arg Lys Ala Met Leu Gln Asp Met Ala IleLeu Thr 65 70 75 80 Gly Gly Gln Val Val Ser Glu Arg Val Gly Leu Ser LeuGlu Thr Ala 85 90 95 Asp Val Ser Leu Leu Gly Gln Ala Arg Lys Val Val ValThr Lys Asp 100 105 110 <210> SEQ ID NO 79 <211> LENGTH: 360 <212> TYPE:DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 79 ccgtacgagaagatcggcgc tgagctggtc aaagaggtcg ccaagaagac cgacgacgtc 60 gcgggcgacggcaccaccac cgccaccgtg ctcgctcagg ctctggttcg cgaaggcctg 120 cgcaacgtcgcagccggcgc caacccgctc ggcctcaagc gtggcatcga gaaggctgtc 180 gaggctgtcacccagtcgct gctgaagtcg gccaaggagg tcgagaccaa ggagcagatt 240 tctgccaccgcggcgatctc cgccggcgac acccagatcg gcgagctcat cgccgaggcc 300 atggacaaggtcggcaacga gggtgtcatc accgtcgagg agtcgaacac cttcggcctg 360 <210> SEQ IDNO 80 <211> LENGTH: 120 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 80 Pro Tyr Glu Lys Ile Gly Ala Glu Leu Val LysGlu Val Ala Lys Lys 1 5 10 15 Thr Asp Asp Val Ala Gly Asp Gly Thr ThrThr Ala Thr Val Leu Ala 20 25 30 Gln Ala Leu Val Arg Glu Gly Leu Arg AsnVal Ala Ala Gly Ala Asn 35 40 45 Pro Leu Gly Leu Lys Arg Gly Ile Glu LysAla Val Glu Ala Val Thr 50 55 60 Gln Ser Leu Leu Lys Ser Ala Lys Glu ValGlu Thr Lys Glu Gln Ile 65 70 75 80 Ser Ala Thr Ala Ala Ile Ser Ala GlyAsp Thr Gln Ile Gly Glu Leu 85 90 95 Ile Ala Glu Ala Met Asp Lys Val GlyAsn Glu Gly Val Ile Thr Val 100 105 110 Glu Glu Ser Asn Thr Phe Gly Leu115 120 <210> SEQ ID NO 81 <211> LENGTH: 43 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Made in a lab <400> SEQUENCE: 81 actgacgctg aggagcgaaa gcgtggggagcgaacaggat tag 43 <210> SEQ ID NO 82 <211> LENGTH: 43 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 82 cgacaaggaa cttcgctaccttaggaccgt catagttacg ggc 43 <210> SEQ ID NO 83 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 83 aaaaaaaaaa aaaaaaaaaa 20<210> SEQ ID NO 84 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 84 ggaaggaagc ggccgctttt tttttttttt t 31 <210> SEQID NO 85 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Made in a lab <400>SEQUENCE: 85 gagagagagc ccgggcatgc tsctsctsct s 31 <210> SEQ ID NO 86<211> LENGTH: 238 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 86 ctcgatgaac cgctcggagc gctcgacctg aagctgcgccacgtcatgca gttcgagctc 60 aagcgcatcc agcgggaggt cgggatcacg ttcatctacgtgacccacga ccaggaagag 120 gcgctcacga tgagtgaccg catcgcggtg atgaacgccggcaacgtcga acagatcggc 180 agcccgaccg agatctacga ccgtcccgcg acggtgttcgtcgccagctt catcgaat 238 <210> SEQ ID NO 87 <211> LENGTH: 79 <212> TYPE:PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 87 Leu Asp GluPro Leu Gly Ala Leu Asp Leu Lys Leu Arg His Val Met 1 5 10 15 Gln PheGlu Leu Lys Arg Ile Gln Arg Glu Val Gly Ile Thr Phe Ile 20 25 30 Tyr ValThr His Asp Gln Glu Glu Ala Leu Thr Met Ser Asp Arg Ile 35 40 45 Ala ValMet Asn Ala Gly Asn Val Glu Gln Ile Gly Ser Pro Thr Glu 50 55 60 Ile TyrAsp Arg Pro Ala Thr Val Phe Val Ala Ser Phe Ile Glu 65 70 75 <210> SEQID NO 88 <211> LENGTH: 1518 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 88 cactcgccat gggtgttaca ataccccaccagttcctcga agtaaacgaa cagaaccgtg 60 acatccagct gagaaaatat tcacagcgacgaagcccggc cgatgcctga tggggtccgg 120 catcagtaca gcgcgctttc ctgcgcggattctattgtcg agtccggggt gtgacgaagg 180 aatccattgt cgaaatgtaa attcgttgcggaatcacttg cataggtccg tcagatccgc 240 gaaggtttac cccacagcca cgacggctgtccccgaggag gacctgccct gaccggcaca 300 cacatcaccg ctgcagaacc tgcagaacagacggcggatt ccgcggcacc gcccaagggc 360 gcgccggtga tcgagatcga ccatgtcacgaagcgcttcg gcgactacct ggccgtcgcg 420 gacgcagact tctccatcgc gcccggggagttcttctcca tgctcggccc gtccgggtgt 480 gggaagacga ccacgttgcg catgatcgcgggattcgaga ccccgactga aggggcgatc 540 cgcctcgaag gcgccgacgt gtcgaggaccccacccaaca agcgcaacgt caacacggtg 600 ttccagcact acgcgctgtt cccgcacatgacggtctggg acaacgtcgc gtacggcccg 660 cgcagcaaga aactcggcaa aggcgaggtccgcaagcgcg tcgacgagct gctggagatc 720 gtccggctga ccgaatttgc cgagcgcaggcccgcccagc tgtccggcgg gcagcagcag 780 cgggtggcgt tggcccgggc actggtgaactaccccagcg cgctgctgct cgatgaaccg 840 ctcggagcgc tcgacctgaa gctgcgccacgtcatgcagt tcgagctcaa gcgcatccag 900 cgggaggtcg ggatcacgtt catctacgtgacccacgacc aggaagaggc gctcacgatg 960 agtgaccgca tcgcggtgat gaacgccggcaacgtcgaac agatcggcag cccgaccgag 1020 atctacgacc gtcccgcgac ggtgttcgtcgccagcttca tcggacaggc caacctctgg 1080 gcgggccggt gcaccggccg ctccaaccgcgattacgtcg agatcgacgt tctcggctcg 1140 acgctgaagg cacgcccggg cgagaccacgatcgagcccg gcgggcacgc caccctgatg 1200 gtgcgtccgg aacgcatccg ggtcaccccgggctcccagg acgcgccgac cggtgacgtc 1260 gcctgcgtgc gtgccaccgt caccgacctgaccttccaag gtccggtggt gcggctctcg 1320 ctggccgctc cggacgactc gaccgtgatcgcccacgtcg gccccgagca ggatctgccg 1380 ctgctgcgcc ccggcgacga cgtgtacgtcagctgggcac cggaagcctc cctggtgctt 1440 cccggcgacg acatccccac caccgaggacctcgaagaga tgctcgacga ctcctgagtc 1500 acgcttcccg attgccga 1518 <210> SEQID NO 89 <211> LENGTH: 376 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 89 Val Ile Glu Ile Asp His Val Thr Lys Arg PheGly Asp Tyr Leu Ala 1 5 10 15 Val Ala Asp Ala Asp Phe Ser Ile Ala ProGly Glu Phe Phe Ser Met 20 25 30 Leu Gly Pro Ser Gly Cys Gly Lys Thr ThrThr Leu Arg Met Ile Ala 35 40 45 Gly Phe Glu Thr Pro Thr Glu Gly Ala IleArg Leu Glu Gly Ala Asp 50 55 60 Val Ser Arg Thr Pro Pro Asn Lys Arg AsnVal Asn Thr Val Phe Gln 65 70 75 80 His Tyr Ala Leu Phe Pro His Met ThrVal Trp Asp Asn Val Ala Tyr 85 90 95 Gly Pro Arg Ser Lys Lys Leu Gly LysGly Glu Val Arg Lys Arg Val 100 105 110 Asp Glu Leu Leu Glu Ile Val ArgLeu Thr Glu Phe Ala Glu Arg Arg 115 120 125 Pro Ala Gln Leu Ser Gly GlyGln Gln Gln Arg Val Ala Leu Ala Arg 130 135 140 Ala Leu Val Asn Tyr ProSer Ala Leu Leu Leu Asp Glu Pro Leu Gly 145 150 155 160 Ala Leu Asp LeuLys Leu Arg His Val Met Gln Phe Glu Leu Lys Arg 165 170 175 Ile Gln ArgGlu Val Gly Ile Thr Phe Ile Tyr Val Thr His Asp Gln 180 185 190 Glu GluAla Leu Thr Met Ser Asp Arg Ile Ala Val Met Asn Ala Gly 195 200 205 AsnVal Glu Gln Ile Gly Ser Pro Thr Glu Ile Tyr Asp Arg Pro Ala 210 215 220Thr Val Phe Val Ala Ser Phe Ile Gly Gln Ala Asn Leu Trp Ala Gly 225 230235 240 Arg Cys Thr Gly Arg Ser Asn Arg Asp Tyr Val Glu Ile Asp Val Leu245 250 255 Gly Ser Thr Leu Lys Ala Arg Pro Gly Glu Thr Thr Ile Glu ProGly 260 265 270 Gly His Ala Thr Leu Met Val Arg Pro Glu Arg Ile Arg ValThr Pro 275 280 285 Gly Ser Gln Asp Ala Pro Thr Gly Asp Val Ala Cys ValArg Ala Thr 290 295 300 Val Thr Asp Leu Thr Phe Gln Gly Pro Val Val ArgLeu Ser Leu Ala 305 310 315 320 Ala Pro Asp Asp Ser Thr Val Ile Ala HisVal Gly Pro Glu Gln Asp 325 330 335 Leu Pro Leu Leu Arg Pro Gly Asp AspVal Tyr Val Ser Trp Ala Pro 340 345 350 Glu Ala Ser Leu Val Leu Pro GlyAsp Asp Ile Pro Thr Thr Glu Asp 355 360 365 Leu Glu Glu Met Leu Asp AspSer 370 375 <210> SEQ ID NO 90 <211> LENGTH: 33 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Made in a lab <400> SEQUENCE: 90 gagagactcg aggtgatcga gatcgaccat gtc 33<210> SEQ ID NO 91 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 91 agagactcga gcaatcggga agcgtgactc a 31 <210> SEQID NO 92 <211> LENGTH: 323 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 92 gtcgactaca aagaagactt caacgacaac gagcagtggttcgccaaggt caaggagccg 60 ttgtcgcgca agcaggacat aggcgccgac ctggtgatccccaccgagtt catggccgcg 120 cgcgtcaagg gcctgggatg gctcaatgag atcagcgaagccggcgtgcc caatcgcaag 180 aatctgcgtc aggacctgtt ggactcgagc atcgacgagggccgcaagtt caccgcgccg 240 tacatgaccg gcatggtcgg tctcgcctac aacaaggcagccaccggacg cgatatccgc 300 accatcgacg acctctggga tcc 323 <210> SEQ ID NO93 <211> LENGTH: 1341 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 93 ccccaccccc ttccctggag ccgacgaaag gcacccgcacatgtcccgtg acatcgatcc 60 ccacctgctg gcccgaatga ccgcacgccg caccttgcgtcgccgcttca tcggcggtgg 120 cgccgcggcc gccgcgggcc tgaccctcgg ttcgtcgttcctggcggcgt gcgggtccga 180 cagtgggacc tcgagcacca cgtcacagga cagcggccccgccagcggcg ccctgcgcgt 240 ctccaactgg ccgctctata tggccgacgg tttcatcgcagcgttccaga ccgcctcggg 300 catcacggtc gactacaaag aagacttcaa cgacaacgagcagtggttcg ccaaggtcaa 360 ggagccgttg tcgcgcaagc aggacatagg cgccgacctggtgatcccca ccgagttcat 420 ggccgcgcgc gtcaagggcc tgggatggct caatgagatcagcgaagccg gcgtgcccaa 480 tcgcaagaat ctgcgtcagg acctgttgga ctcgagcatcgacgagggcc gcaagttcac 540 cgcgccgtac atgaccggca tggtcggtct cgcctacaacaaggcagcca ccggacgcga 600 tatccgcacc atcgacgacc tctgggatcc cgcgttcaagggccgcgtca gtctgttctc 660 cgacgtccag gacggcctcg gcatgatcat gctctcgcagggcaactcgc cggagaatcc 720 gaccaccgag tccattcagc aggcggtcga tctggtccgcgaacagaacg acagggggtc 780 agatccgtcg cttcaccggc aacgactacg ccgacgacctggccgcagaa acatcgccat 840 cgcgcaggcg tactccggtg acgtcgtgca gctgcaggcggacaaccccg atctgcagtt 900 catcgttccc gaatccggcg gcgactggtt cgtcgacacgatggtgatcc cgtacaccac 960 gcagaaccag aaggccgccg aggcgtggat cgactacatctacgaccgag ccaactacgc 1020 caagctggtc gcgttcaccc agttcgtgcc cgcactctcggacatgaccg acgaactcgc 1080 caaggtcgat cctgcatcgg cggagaaccc gctgatcaacccgtcggccg aggtgcaggc 1140 gaacctgaag tcgtgggcgg cactgaccga cgagcagacgcaggagttca acactgcgta 1200 cgccgccgtc accggcggct gacgcggtgg tagtgccgatgcgaggggca taaatggccc 1260 tgcggacgcg aggagcataa atggccggtg tcgccaccagcagccgtcag cggacaaggt 1320 cgctccgtat ctgatggtcc t 1341 <210> SEQ ID NO94 <211> LENGTH: 393 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 94 Met Ser Arg Asp Ile Asp Pro His Leu Leu AlaArg Met Thr Ala Arg 1 5 10 15 Arg Thr Leu Arg Arg Arg Phe Ile Gly GlyGly Ala Ala Ala Ala Ala 20 25 30 Gly Leu Thr Leu Gly Ser Ser Phe Leu AlaAla Cys Gly Ser Asp Ser 35 40 45 Gly Thr Ser Ser Thr Thr Ser Gln Asp SerGly Pro Ala Ser Gly Ala 50 55 60 Leu Arg Val Ser Asn Trp Pro Leu Tyr MetAla Asp Gly Phe Ile Ala 65 70 75 80 Ala Phe Gln Thr Ala Ser Gly Ile ThrVal Asp Tyr Lys Glu Asp Phe 85 90 95 Asn Asp Asn Glu Gln Trp Phe Ala LysVal Lys Glu Pro Leu Ser Arg 100 105 110 Lys Gln Asp Ile Gly Ala Asp LeuVal Ile Pro Thr Glu Phe Met Ala 115 120 125 Ala Arg Val Lys Gly Leu GlyTrp Leu Asn Glu Ile Ser Glu Ala Gly 130 135 140 Val Pro Asn Arg Lys AsnLeu Arg Gln Asp Leu Leu Asp Ser Ser Ile 145 150 155 160 Asp Glu Gly ArgLys Phe Thr Ala Pro Tyr Met Thr Gly Met Val Gly 165 170 175 Leu Ala TyrAsn Lys Ala Ala Thr Gly Arg Asp Ile Arg Thr Ile Asp 180 185 190 Asp LeuTrp Asp Pro Ala Phe Lys Gly Arg Val Ser Leu Phe Ser Asp 195 200 205 ValGln Asp Gly Leu Gly Met Ile Met Leu Ser Gln Gly Asn Ser Pro 210 215 220Glu Asn Pro Thr Thr Glu Ser Ile Gln Gln Ala Val Asp Leu Val Arg 225 230235 240 Glu Gln Asn Asp Arg Gly Ser Asp Pro Ser Leu His Arg Gln Arg Leu245 250 255 Arg Arg Arg Pro Gly Arg Arg Asn Ile Ala Ile Ala Gln Ala TyrSer 260 265 270 Gly Asp Val Val Gln Leu Gln Ala Asp Asn Pro Asp Leu GlnPhe Ile 275 280 285 Val Pro Glu Ser Gly Gly Asp Trp Phe Val Asp Thr MetVal Ile Pro 290 295 300 Tyr Thr Thr Gln Asn Gln Lys Ala Ala Glu Ala TrpIle Asp Tyr Ile 305 310 315 320 Tyr Asp Arg Ala Asn Tyr Ala Lys Leu ValAla Phe Thr Gln Phe Val 325 330 335 Pro Ala Leu Ser Asp Met Thr Asp GluLeu Ala Lys Val Asp Pro Ala 340 345 350 Ser Ala Glu Asn Pro Leu Ile AsnPro Ser Ala Glu Val Gln Ala Asn 355 360 365 Leu Lys Ser Trp Ala Ala LeuThr Asp Glu Gln Thr Gln Glu Phe Asn 370 375 380 Thr Ala Tyr Ala Ala ValThr Gly Gly 385 390 <210> SEQ ID NO 95 <211> LENGTH: 22 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 95 atgtcccgtgacatcgatcc cc 22 <210> SEQ ID NO 96 <211> LENGTH: 21 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 96 atcggcactaccaccgcgtc a 21 <210> SEQ ID NO 97 <211> LENGTH: 861 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 97 gccggcgctcgcatatctcg cgatcttctt ccgtggtgcc gttcttctcg ctggcacgca 60 cctcgttgtcggagaccggc ggctcggtgt tcatgccgac gctgacgttc gcctgggact 120 tcggcaactacgtcgacgcg ttcacgatgt accacgagca gatcttccgc tcgttcggct 180 acgcgttcgtcgccacggtg ctgtgcctgt tgctggcgtt cccgctggcc tacgtcatcg 240 cgttcaaggccggccggttc aagaacctga tcctggggct ggtgatcctg ccgttcttcg 300 tcacgttcctgatccgcacc attgcgtgga agacgatcct ggccgacgaa ggctgggtgg 360 tcaccgcgctgggcgccatc gggctgctgc ctgacgaggg ccggctgctg tccaccagct 420 gggcggtcatcggcggtctg acctacaact ggatcatctt catgatcctg ccgctgtacg 480 tcagcctggagaagatcgac ccgcgtctgc tggaggcctc ccaggacctc tactcgtcgg 540 cgccgcgcagcttcggcaag gtgatcctgc cgatggcgat gcccggggtg ctggccggga 600 gcatgctggtgttcatcccg gccgtcggcg acttcatcaa cgccgactat ctcggcagta 660 cccagaccaccatgatcggc aacgtgatcc agaagcagtt cctggtcgtc aaggactatc 720 cggcggcggccgcgctgagt ctggggctga tgttgctgat cctgatcggc gtgctcctct 780 acacacgggcgctgggttcg gaggatctgg tatgaccacc caggcaggcg ccgcactggc 840 caccgccgcccagcaggatc c 861 <210> SEQ ID NO 98 <211> LENGTH: 259 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 98 Val Val Pro PhePhe Ser Leu Ala Arg Thr Ser Leu Ser Glu Thr Gly 1 5 10 15 Gly Ser ValPhe Met Pro Thr Leu Thr Phe Ala Trp Asp Phe Gly Asn 20 25 30 Tyr Val AspAla Phe Thr Met Tyr His Glu Gln Ile Phe Arg Ser Phe 35 40 45 Gly Tyr AlaPhe Val Ala Thr Val Leu Cys Leu Leu Leu Ala Phe Pro 50 55 60 Leu Ala TyrVal Ile Ala Phe Lys Ala Gly Arg Phe Lys Asn Leu Ile 65 70 75 80 Leu GlyLeu Val Ile Leu Pro Phe Phe Val Thr Phe Leu Ile Arg Thr 85 90 95 Ile AlaTrp Thr Ile Leu Ala Asp Glu Gly Trp Val Val Thr Ala Leu 100 105 110 GlyAla Ile Gly Leu Leu Pro Asp Glu Gly Arg Leu Leu Ser Thr Ser 115 120 125Trp Ala Val Ile Gly Gly Leu Thr Tyr Asn Trp Ile Ile Phe Met Ile 130 135140 Leu Pro Leu Tyr Val Ser Leu Glu Lys Ile Asp Pro Arg Leu Leu Glu 145150 155 160 Ala Ser Gln Asp Leu Tyr Ser Ser Ala Pro Arg Ser Phe Gly LysVal 165 170 175 Ile Leu Pro Met Ala Met Pro Gly Val Leu Ala Gly Ser MetLeu Val 180 185 190 Phe Ile Pro Ala Val Gly Asp Phe Ile Asn Ala Asp TyrLeu Gly Ser 195 200 205 Thr Gln Thr Thr Met Ile Gly Asn Val Ile Gln LysGln Phe Leu Val 210 215 220 Val Lys Asp Tyr Pro Ala Ala Ala Ala Leu SerLeu Gly Leu Met Leu 225 230 235 240 Leu Ile Leu Ile Gly Val Leu Leu TyrThr Arg Ala Leu Gly Ser Glu 245 250 255 Asp Leu Val <210> SEQ ID NO 99<211> LENGTH: 277 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 99 gtaatctttg ctggagcccg tacgccggta ggcaaactcatgggttcgct caaggacttc 60 aagggcagcg atctcggtgc cgtggcgatc aagggcgccctggagaaagc cttccccggc 120 gtcgacgacc ctgctcgtct cgtcgagtac gtgatcatgggccaagtgct ctccgccggc 180 gccggccaga tgcccgcccg ccaggccgcc gtcgccgccggcatcccgtg ggacgtcgcc 240 tcgctgacga tcaacaagat gtgcctgtcg ggcatcg 277<210> SEQ ID NO 100 <211> LENGTH: 92 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 100 Val Ile Phe Ala Gly Ala Arg ThrPro Val Gly Lys Leu Met Gly Ser 1 5 10 15 Leu Lys Asp Phe Lys Gly SerAsp Leu Gly Ala Val Ala Ile Lys Gly 20 25 30 Ala Leu Glu Lys Ala Phe ProGly Val Asp Asp Pro Ala Arg Leu Val 35 40 45 Glu Tyr Val Ile Met Gly GlnVal Leu Ser Ala Gly Ala Gly Gln Met 50 55 60 Pro Ala Arg Gln Ala Ala ValAla Ala Gly Ile Pro Trp Asp Val Ala 65 70 75 80 Ser Leu Thr Ile Asn LysMet Cys Leu Ser Gly Ile 85 90 <210> SEQ ID NO 101 <211> LENGTH: 12 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (1)...(1) <223> OTHER INFORMATION:Residue can be either Glu or Pro <221> NAME/KEY: UNSURE <222> LOCATION:(2)...(2) <223> OTHER INFORMATION: Residue can be either Pro or Glu<221> NAME/KEY: UNSURE <222> LOCATION: (7)...(7) <221> NAME/KEY: UNSURE<222> LOCATION: (12)...(12) <400> SEQUENCE: 101 Xaa Xaa Ala Asp Arg GlyXaa Ser Lys Tyr Arg Xaa 1 5 10 <210> SEQ ID NO 102 <211> LENGTH: 24<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (1)...(1) <400> SEQUENCE: 102 XaaIle Asp Glu Ser Leu Phe Asp Ala Glu Glu Lys Met Glu Lys Ala 1 5 10 15Val Ser Val Ala Arg Asp Ser Ala 20 <210> SEQ ID NO 103 <211> LENGTH: 23<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (1)...(2) <221> NAME/KEY: UNSURE<222> LOCATION: (15)...(15) <221> NAME/KEY: UNSURE <222> LOCATION:(17)...(17) <400> SEQUENCE: 103 Xaa Xaa Ile Ala Pro Ala Thr Ser Gly ThrLeu Ser Glu Phe Xaa Ala 1 5 10 15 Xaa Lys Gly Val Thr Met Glu 20 <210>SEQ ID NO 104 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 104 Pro Asn Val Pro Asp Ala Phe AlaVal Leu Ala Asp Arg Val Gly 1 5 10 15 <210> SEQ ID NO 105 <211> LENGTH:9 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (1)...(1) <400> SEQUENCE: 105 XaaIle Arg Val Gly Val Asn Gly Phe 1 5 <210> SEQ ID NO 106 <211> LENGTH:485 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE:106 agcggctggg acatcaacac cgccgccttc gagtggtacg tcgactcggg tctcgcggtg 60atcatgcccg tcggcgggca gtccagcttc tacagcgact ggtacagccc ggcctgcggt 120aaggccggct gccagaccta caagtgggag acgttcctga cccaggagct gccggcctac 180ctcgccgcca acaagggggt cgacccgaac cgcaacgcgg ccgtcggtct gtccatggcc 240ggttcggcgg cgctgacgct ggcgatctac cacccgcagc agttccagta cgccgggtcg 300ctgtcgggct acctgaaccc gtccgagggg tggtggccga tgctgatcaa catctcgatg 360ggtgacgcgg gcggctacaa ggccaacgac atgtggggtc gcaccgagga cccgagcagc 420gcctggaagc gcaacgaccc gatggtcaac atcggcaagc tggtcgccaa caacaccccc 480ctctc 485 <210> SEQ ID NO 107 <211> LENGTH: 501 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: unsure<222> LOCATION: (441)...(441) <221> NAME/KEY: unsure <222> LOCATION:(450)...(450) <400> SEQUENCE: 107 atgccggtgc gacgtgcgcg cagtgcgcttgcgtccgtga ccttcgtcgc ggccgcgtgc 60 gtgggcgctg agggcaccgc actggcggcgacgccggact ggagcgggcg ctacacggtg 120 gtgacgttcg cctccgacaa actcggcacgagtgtggccg cccgccagcc agaacccgac 180 ttcagcggtc agtacacctt cagcacgtcctgtgtgggca cctgcgtggc caccgcgtcc 240 gacggcccgg cgccgtcgaa cccgacgattccgcagcccg cgcgctacac ctgggacggc 300 aggcagtggg tgttcaacta caactggcagtgggagtgct tccgcggcgc cgacgtcccg 360 cgcgagtacg ccgccgcgcg ttcgctggtgttctacgccc cgaccgccga cgggtcgatg 420 ttcggcacct ggcgcaccga natcctgganggcctctgca agggcaccgt gatcatgccg 480 gtcgcggcct atccggcgta g 501 <210>SEQ ID NO 108 <211> LENGTH: 180 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 108 atgaaccagc cgcggcccgaggccgaggcg aacctgcggg gctacttcac cgccaacccg 60 gcggagtact acgacctgcggggcatcctc gccccgatcg gtgacgcgca gcgcaactgc 120 aacatcaccg tgctgccggtagagctgcag acggcctacg acacgttcat ggccggctga 180 <210> SEQ ID NO 109<211> LENGTH: 166 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 109 Met Pro Val Arg Arg Ala Arg Ser Ala Leu Ala Ser ValThr Phe Val 1 5 10 15 Ala Ala Ala Cys Val Gly Ala Glu Gly Thr Ala LeuAla Ala Thr Pro 20 25 30 Asp Trp Ser Gly Arg Tyr Thr Val Val Thr Phe AlaSer Asp Lys Leu 35 40 45 Gly Thr Ser Val Ala Ala Arg Gln Pro Glu Pro AspPhe Ser Gly Gln 50 55 60 Tyr Thr Phe Ser Thr Ser Cys Val Gly Thr Cys ValAla Thr Ala Ser 65 70 75 80 Asp Gly Pro Ala Pro Ser Asn Pro Thr Ile ProGln Pro Ala Arg Tyr 85 90 95 Thr Trp Asp Gly Arg Gln Trp Val Phe Asn TyrAsn Trp Gln Trp Glu 100 105 110 Cys Phe Arg Gly Ala Asp Val Pro Arg GluTyr Ala Ala Ala Arg Ser 115 120 125 Leu Val Phe Tyr Ala Pro Thr Ala AspGly Ser Met Phe Gly Thr Trp 130 135 140 Arg Thr Asp Ile Leu Asp Gly LeuCys Lys Gly Thr Val Ile Met Pro 145 150 155 160 Val Ala Ala Tyr Pro Ala165 <210> SEQ ID NO 110 <211> LENGTH: 74 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 110 Pro Arg Asp Thr His Pro Gly AlaAsn Gln Ala Val Thr Ala Ala Met 1 5 10 15 Asn Gln Pro Arg Pro Glu AlaGlu Ala Asn Leu Arg Gly Tyr Phe Thr 20 25 30 Ala Asn Pro Ala Glu Tyr TyrAsp Leu Arg Gly Ile Leu Ala Pro Ile 35 40 45 Gly Asp Ala Gln Arg Asn CysAsn Ile Thr Val Leu Pro Val Glu Leu 50 55 60 Gln Thr Ala Tyr Asp Thr PheMet Ala Gly 65 70 <210> SEQ ID NO 111 <211> LENGTH: 503 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (358)...(358) <400> SEQUENCE: 111 atgcaggtgcggcgtgttct gggcagtgtc ggtgcagcag tcgcggtttc ggccgcgtta 60 tggcagacgggggtttcgat accgaccgcc tcagcggatc cgtgtccgga catcgaggtg 120 atcttcgcgcgcgggaccgg tgcggaaccc ggcctcgggt gggtcggtga tgcgttcgtc 180 aacgcgctgcggcccaaggt cggtgagcag tcggtgggca cctacgcggt gaactacccg 240 gcaggattcggacttcgaca aatcggcgcc catgggcgcg gccgacgcat cggggcgggt 300 gcagtggatggccgacaact gcccggacac caagcttgtc ctgggcggca tgtcgcangg 360 cgccggcgtcatcgacctga tcaccgtcga tccgcgaccg ctgggccggt tcacccccac 420 cccgatgccgccccgcgtcg ccgaccacgt ggccgccgtt gtggtcttcg gaaatccgtt 480 gcgcgacatccgtggtggcg gtc 503 <210> SEQ ID NO 112 <211> LENGTH: 167 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (119)...(119) <400> SEQUENCE: 112 Met Gln Val ArgArg Val Leu Gly Ser Val Gly Ala Ala Val Ala Val 1 5 10 15 Ser Ala AlaLeu Trp Gln Thr Gly Val Ser Ile Pro Thr Ala Ser Ala 20 25 30 Asp Pro CysPro Asp Ile Glu Val Ile Phe Ala Arg Gly Thr Gly Ala 35 40 45 Glu Pro GlyLeu Gly Trp Val Gly Asp Ala Phe Val Asn Ala Leu Arg 50 55 60 Pro Lys ValGly Glu Gln Ser Val Gly Thr Tyr Ala Val Asn Tyr Pro 65 70 75 80 Ala GlyPhe Asp Phe Asp Lys Ser Ala Pro Met Gly Ala Ala Asp Ala 85 90 95 Ser GlyArg Val Gln Trp Met Ala Asp Asn Cys Pro Asp Thr Lys Leu 100 105 110 ValLeu Gly Gly Met Ser Xaa Gly Ala Gly Val Ile Asp Leu Ile Thr 115 120 125Val Asp Pro Arg Pro Leu Gly Arg Phe Thr Pro Thr Pro Met Pro Pro 130 135140 Arg Val Ala Asp His Val Ala Ala Val Val Val Phe Gly Asn Pro Leu 145150 155 160 Arg Asp Ile Arg Gly Gly Gly 165 <210> SEQ ID NO 113 <211>LENGTH: 1569 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 113 atggccaaga caattgcgta tgacgaagag gcccgccgtg gcctcgagcggggcctcaac 60 gccctcgcag acgccgtaaa ggtgacgttg ggcccgaagg gtcgcaacgtcgtgctggag 120 aagaagtggg gcgcccccac gatcaccaac gatggtgtgt ccatcgccaaggagatcgag 180 ctggaggacc cgtacgagaa gatcggcgct gagctggtca aagaggtcgccaagaagacc 240 gacgacgtcg cgggcgacgg caccaccacc gccaccgtgc tcgctcaggctctggttcgc 300 gaaggcctgc gcaacgtcgc agccggcgcc aacccgctcg gcctcaagcgtggcatcgag 360 aaggctgtcg aggctgtcac ccagtcgctg ctgaagtcgg ccaaggaggtcgagaccaag 420 gagcagattt ctgccaccgc ggcgatttcc gccggcgaca cccagatcggcgagctcatc 480 gccgaggcca tggacaaggt cggcaacgag ggtgtcatca ccgtcgaggagtcgaacacc 540 ttcggcctgc agctcgagct caccgagggt atgcgcttcg acaagggctacatctcgggt 600 tacttcgtga ccgacgccga gcgccaggaa gccgtcctgg aggatccctacatcctgctg 660 gtcagctcca aggtgtcgac cgtcaaggat ctgctcccgc tgctggagaaggtcatccag 720 gccggcaagc cgctgctgat catcgccgag gacgtcgagg gcgaggccctgtccacgctg 780 gtggtcaaca agatccgcgg caccttcaag tccgtcgccg tcaaggctccgggcttcggt 840 gaccgccgca aggcgatgct gcaggacatg gccatcctca ccggtggtcaggtcgtcagc 900 gaaagagtcg ggctgtccct ggagaccgcc gacgtctcgc tgctgggccaggcccgcaag 960 gtcgtcgtca ccaaggacga gaccaccatc gtcgagggct cgggcgattccgatgccatc 1020 gccggccggg tggctcagat ccgcgccgag atcgagaaca gcgactccgactacgaccgc 1080 gagaagctgc aggagcgcct ggccaagctg gccggcggtg ttgcggtgatcaaggccgga 1140 gctgccaccg aggtggagct caaggagcgc aagcaccgca tcgaggacgccgtccgcaac 1200 gcgaaggctg ccgtcgaaga gggcatcgtc gccggtggcg gcgtggctctgctgcagtcg 1260 gctcctgcgc tggacgacct cggcctgacg ggcgacgagg ccaccggtgccaacatcgtc 1320 cgcgtggcgc tgtcggctcc gctcaagcag atcgccttca acggcggcctggagcccggc 1380 gtcgttgccg agaaggtgtc caacctgccc gcgggtcacg gcctcaacgccgcgaccggt 1440 gagtacgagg acctgctcaa ggccggcgtc gccgacccgg tgaaggtcacccgctcggcg 1500 ctgcagaacg cggcgtccat cgcggctctg ttcctcacca ccgaggccgtcgtcgccgac 1560 aagccggag 1569 <210> SEQ ID NO 114 <211> LENGTH: 523<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 114Met Ala Lys Thr Ile Ala Tyr Asp Glu Glu Ala Arg Arg Gly Leu Glu 1 5 1015 Arg Gly Leu Asn Ala Leu Ala Asp Ala Val Lys Val Thr Leu Gly Pro 20 2530 Lys Gly Arg Asn Val Val Leu Glu Lys Lys Trp Gly Ala Pro Thr Ile 35 4045 Thr Asn Asp Gly Val Ser Ile Ala Lys Glu Ile Glu Leu Glu Asp Pro 50 5560 Tyr Glu Lys Ile Gly Ala Glu Leu Val Lys Glu Val Ala Lys Lys Thr 65 7075 80 Asp Asp Val Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala Gln 8590 95 Ala Leu Val Arg Glu Gly Leu Arg Asn Val Ala Ala Gly Ala Asn Pro100 105 110 Leu Gly Leu Lys Arg Gly Ile Glu Lys Ala Val Glu Ala Val ThrGln 115 120 125 Ser Leu Leu Lys Ser Ala Lys Glu Val Glu Thr Lys Glu GlnIle Ser 130 135 140 Ala Thr Ala Ala Ile Ser Ala Gly Asp Thr Gln Ile GlyGlu Leu Ile 145 150 155 160 Ala Glu Ala Met Asp Lys Val Gly Asn Glu GlyVal Ile Thr Val Glu 165 170 175 Glu Ser Asn Thr Phe Gly Leu Gln Leu GluLeu Thr Glu Gly Met Arg 180 185 190 Phe Asp Lys Gly Tyr Ile Ser Gly TyrPhe Val Thr Asp Ala Glu Arg 195 200 205 Gln Glu Ala Val Leu Glu Asp ProTyr Ile Leu Leu Val Ser Ser Lys 210 215 220 Val Ser Thr Val Lys Asp LeuLeu Pro Leu Leu Glu Lys Val Ile Gln 225 230 235 240 Ala Gly Lys Pro LeuLeu Ile Ile Ala Glu Asp Val Glu Gly Glu Ala 245 250 255 Leu Ser Thr LeuVal Val Asn Lys Ile Arg Gly Thr Phe Lys Ser Val 260 265 270 Ala Val LysAla Pro Gly Phe Gly Asp Arg Arg Lys Ala Met Leu Gln 275 280 285 Asp MetAla Ile Leu Thr Gly Gly Gln Val Val Ser Glu Arg Val Gly 290 295 300 LeuSer Leu Glu Thr Ala Asp Val Ser Leu Leu Gly Gln Ala Arg Lys 305 310 315320 Val Val Val Thr Lys Asp Glu Thr Thr Ile Val Glu Gly Ser Gly Asp 325330 335 Ser Asp Ala Ile Ala Gly Arg Val Ala Gln Ile Arg Ala Glu Ile Glu340 345 350 Asn Ser Asp Ser Asp Tyr Asp Arg Glu Lys Leu Gln Glu Arg LeuAla 355 360 365 Lys Leu Ala Gly Gly Val Ala Val Ile Lys Ala Gly Ala AlaThr Glu 370 375 380 Val Glu Leu Lys Glu Arg Lys His Arg Ile Glu Asp AlaVal Arg Asn 385 390 395 400 Ala Lys Ala Ala Val Glu Glu Gly Ile Val AlaGly Gly Gly Val Ala 405 410 415 Leu Leu Gln Ser Ala Pro Ala Leu Asp AspLeu Gly Leu Thr Gly Asp 420 425 430 Glu Ala Thr Gly Ala Asn Ile Val ArgVal Ala Leu Ser Ala Pro Leu 435 440 445 Lys Gln Ile Ala Phe Asn Gly GlyLeu Glu Pro Gly Val Val Ala Glu 450 455 460 Lys Val Ser Asn Leu Pro AlaGly His Gly Leu Asn Ala Ala Thr Gly 465 470 475 480 Glu Tyr Glu Asp LeuLeu Lys Ala Gly Val Ala Asp Pro Val Lys Val 485 490 495 Thr Arg Ser AlaLeu Gln Asn Ala Ala Ser Ile Ala Ala Leu Phe Leu 500 505 510 Thr Thr GluAla Val Val Ala Asp Lys Pro Glu 515 520 <210> SEQ ID NO 115 <211>LENGTH: 647 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 115 atggccaaga caattgcgta tgacgaagag gcccgccgtg gcctcgagcggggcctcaac 60 gccctcgcag acgccgtaaa ggtgacgttg ggcccgaagg gtcgcaacgtcgtgctggag 120 aagaagtggg gcgcccccac gatcaccaac gatggtgtgt ccatcgccaaggagatcgag 180 ctggaggacc cgtacgagaa gatcggcgct gagctggtca aagaggtcgccaagaagacc 240 gacgacgtcg cgggcgacgg caccaccacc gccaccgtgc tcgctcaggctctggttcgc 300 gaaggcctgc gcaacgtcgc agccggcgcc aacccgctcg gcctcaagcgtggcatcgag 360 aaggctgtcg aggctgtcac ccagtcgctg ctgaagtcgg ccaaggaggtcgagaccaag 420 gagcagattt ctgccaccgc ggcgatttcc gccggcgaca cccagatcggcgagctcatc 480 gccgaggcca tggacaaggt cggcaacgag ggtgtcatca ccgtcgaggagtcgaacacc 540 ttcggcctgc agctcgagct caccgagggt atgcgcttcg acaagggctacatctcgggt 600 tacttcgtga ccgacgccga gcgccaggaa gccgtcctgg aggatcc 647<210> SEQ ID NO 116 <211> LENGTH: 927 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 116 gatccctaca tcctgctggtcagctccaag gtgtcgaccg tcaaggatct gctcccgctg 60 ctggagaagg tcatccaggccggcaagccg ctgctgatca tcgccgagga cgtcgagggc 120 gaggccctgt ccacgctggtggtcaacaag atccgcggca ccttcaagtc cgtcgccgtc 180 aaggctccgg gcttcggtgaccgccgcaag gcgatgctgc aggacatggc catcctcacc 240 ggtggtcagg tcgtcagcgaaagagtcggg ctgtccctgg agaccgccga cgtctcgctg 300 ctgggccagg cccgcaaggtcgtcgtcacc aaggacgaga ccaccatcgt cgagggctcg 360 ggcgattccg atgccatcgccggccgggtg gctcagatcc gcgccgagat cgagaacagc 420 gactccgact acgaccgcgagaagctgcag gagcgcctgg ccaagctggc cggcggtgtt 480 gcggtgatca aggccggagctgccaccgag gtggagctca aggagcgcaa gcaccgcatc 540 gaggacgccg tccgcaacgcgaaggctgcc gtcgaagagg gcatcgtcgc cggtggcggc 600 gtggctctgc tgcagtcggctcctgcgctg gacgacctcg gcctgacggg cgacgaggcc 660 accggtgcca acatcgtccgcgtggcgctg tcggctccgc tcaagcagat cgccttcaac 720 ggcggcctgg agcccggcgtcgttgccgag aaggtgtcca acctgcccgc gggtcacggc 780 ctcaacgccg cgaccggtgagtacgaggac ctgctcaagg ccggcgtcgc cgacccggtg 840 aaggtcaccc gctcggcgctgcagaacgcg gcgtccatcg cggctctgtt cctcaccacc 900 gaggccgtcg tcgccgacaagccggag 927 <210> SEQ ID NO 117 <211> LENGTH: 215 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 117 Met Ala Lys Thr IleAla Tyr Asp Glu Glu Ala Arg Arg Gly Leu Glu 1 5 10 15 Arg Gly Leu AsnAla Leu Ala Asp Ala Val Lys Val Thr Leu Gly Pro 20 25 30 Lys Gly Arg AsnVal Val Leu Glu Lys Lys Trp Gly Ala Pro Thr Ile 35 40 45 Thr Asn Asp GlyVal Ser Ile Ala Lys Glu Ile Glu Leu Glu Asp Pro 50 55 60 Tyr Glu Lys IleGly Ala Glu Leu Val Lys Glu Val Ala Lys Lys Thr 65 70 75 80 Asp Asp ValAla Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala Gln 85 90 95 Ala Leu ValArg Glu Gly Leu Arg Asn Val Ala Ala Gly Ala Asn Pro 100 105 110 Leu GlyLeu Lys Arg Gly Ile Glu Lys Ala Val Glu Ala Val Thr Gln 115 120 125 SerLeu Leu Lys Ser Ala Lys Glu Val Glu Thr Lys Glu Gln Ile Ser 130 135 140Ala Thr Ala Ala Ile Ser Ala Gly Asp Thr Gln Ile Gly Glu Leu Ile 145 150155 160 Ala Glu Ala Met Asp Lys Val Gly Asn Glu Gly Val Ile Thr Val Glu165 170 175 Glu Ser Asn Thr Phe Gly Leu Gln Leu Glu Leu Thr Glu Gly MetArg 180 185 190 Phe Asp Lys Gly Tyr Ile Ser Gly Tyr Phe Val Thr Asp AlaGlu Arg 195 200 205 Gln Glu Ala Val Leu Glu Asp 210 215 <210> SEQ ID NO118 <211> LENGTH: 309 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 118 Asp Pro Tyr Ile Leu Leu Val Ser Ser Lys ValSer Thr Val Lys Asp 1 5 10 15 Leu Leu Pro Leu Leu Glu Lys Val Ile GlnAla Gly Lys Pro Leu Leu 20 25 30 Ile Ile Ala Glu Asp Val Glu Gly Glu AlaLeu Ser Thr Leu Val Val 35 40 45 Asn Lys Ile Arg Gly Thr Phe Lys Ser ValAla Val Lys Ala Pro Gly 50 55 60 Phe Gly Asp Arg Arg Lys Ala Met Leu GlnAsp Met Ala Ile Leu Thr 65 70 75 80 Gly Gly Gln Val Val Ser Glu Arg ValGly Leu Ser Leu Glu Thr Ala 85 90 95 Asp Val Ser Leu Leu Gly Gln Ala ArgLys Val Val Val Thr Lys Asp 100 105 110 Glu Thr Thr Ile Val Glu Gly SerGly Asp Ser Asp Ala Ile Ala Gly 115 120 125 Arg Val Ala Gln Ile Arg AlaGlu Ile Glu Asn Ser Asp Ser Asp Tyr 130 135 140 Asp Arg Glu Lys Leu GlnGlu Arg Leu Ala Lys Leu Ala Gly Gly Val 145 150 155 160 Ala Val Ile LysAla Gly Ala Ala Thr Glu Val Glu Leu Lys Glu Arg 165 170 175 Lys His ArgIle Glu Asp Ala Val Arg Asn Ala Lys Ala Ala Val Glu 180 185 190 Glu GlyIle Val Ala Gly Gly Gly Val Ala Leu Leu Gln Ser Ala Pro 195 200 205 AlaLeu Asp Asp Leu Gly Leu Thr Gly Asp Glu Ala Thr Gly Ala Asn 210 215 220Ile Val Arg Val Ala Leu Ser Ala Pro Leu Lys Gln Ile Ala Phe Asn 225 230235 240 Gly Gly Leu Glu Pro Gly Val Val Ala Glu Lys Val Ser Asn Leu Pro245 250 255 Ala Gly His Gly Leu Asn Ala Ala Thr Gly Glu Tyr Glu Asp LeuLeu 260 265 270 Lys Ala Gly Val Ala Asp Pro Val Lys Val Thr Arg Ser AlaLeu Gln 275 280 285 Asn Ala Ala Ser Ile Ala Ala Leu Phe Leu Thr Thr GluAla Val Val 290 295 300 Ala Asp Lys Pro Glu 305 <210> SEQ ID NO 119<211> LENGTH: 162 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 119 ctcgtacagg cgacggagat ctccgacgac gccacgtcggtacggttggt cgccaccctg 60 ttcggcgtcg tgttgttgac gttggtgctg tccgggctcaacgccaccct catccagggc 120 gcaccagaag acagctggcg caggcggatt ccgtcgatct tc162 <210> SEQ ID NO 120 <211> LENGTH: 1366 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: unsure<222> LOCATION: (955)...(955) <221> NAME/KEY: unsure <222> LOCATION:(973)...(973) <400> SEQUENCE: 120 gatgagcagc gtgctgaact cgacctggttggcctgggcc gtcgcggtcg cggtcgggtt 60 cccggtgctg ctggtcgtgc tgaccgaggtgcacaacgcg ttgcgtcggc gcggcagcgc 120 gctggcccgc ccggtgcaac tcctgcgtacctacatcctg ccgctgggcg cgttgctgct 180 cctgctggta caggcgatgg agatctccgacgacgccacg tcggtacggt tggtcgccac 240 cctgttcggc gtcgtgttgt tgacgttggtgctgtccggg ctcaacgcca ccctcatcca 300 gggcgcacca gaagacagct ggcgcaggcggattccgtcg atcttcctcg acgtcgcgcg 360 cttcgcgctg atcgcggtcg gtatcaccgtgatcatggcc tatgtctggg gcgcgaacgt 420 ggggggcctg ttcaccgcac tgggcgtcacttccatcgtt cttggcctgg ctctgcagaa 480 ttcggtcggt cagatcatct cgggtctgctgctgctgttc gagcaaccgt tccggctcgg 540 cgactggatc accgtcccca ccgcggcgggccggccgtcc gcccacggcc gcgtggtgga 600 agtcaactgg cgtgcaacac atatcgacaccggcggcaac ctgctggtaa tgcccaacgc 660 cgaactcgcc ggcgcgtcgt tcaccaattacagccggccc gtgggagagc accggctgac 720 cgtcgtcacc accttcaacg ccgcggacacccccgatgat gtctgcgaga tgctgtcgtc 780 ggtcgcggcg tcgctgcccg aactgcgcaccgacggacag atcgccacgc tctatctcgg 840 tgcggccgaa tacgagaagt cgatcccgttgcacacaccc gcggtggacg actcggtcag 900 gagcacgtac ctgcgatggg tctggtacgccgcgcgccgg caggaacttc gcctnaacgg 960 cgtcgccgac ganttcgaca cgccggaacggatcgcctcg gccatgcggg ctgtggcgtc 1020 cacactgcgc ttggcagacg acgaacagcaggagatcgcc gacgtggtgc gtctggtccg 1080 ttacggcaac ggggaacgcc tccagcagccgggtcaggta ccgaccggga tgaggttcat 1140 cgtagacggc agggtgagtc tgtccgtgatcgatcaggac ggcgacgtga tcccggcgcg 1200 ggtgctcgag cgtggcgact tcctggggcagaccacgctg acgcgggaac cggtactggc 1260 gaccgcgcac gcgctggagg aagtcaccgtgctggagatg gcccgtgacg agatcgagcg 1320 cctggtgcac cgaaagccga tcctgctgcacgtgatcggg gccgtg 1366 <210> SEQ ID NO 121 <211> LENGTH: 455 <212> TYPE:PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (318)...(318) <221> NAME/KEY: UNSURE <222>LOCATION: (324)...(324) <400> SEQUENCE: 121 Met Ser Ser Val Leu Asn SerThr Trp Leu Ala Trp Ala Val Ala Val 1 5 10 15 Ala Val Gly Phe Pro ValLeu Leu Val Val Leu Thr Glu Val His Asn 20 25 30 Ala Leu Arg Arg Arg GlySer Ala Leu Ala Arg Pro Val Gln Leu Leu 35 40 45 Arg Thr Tyr Ile Leu ProLeu Gly Ala Leu Leu Leu Leu Leu Val Gln 50 55 60 Ala Met Glu Ile Ser AspAsp Ala Thr Ser Val Arg Leu Val Ala Thr 65 70 75 80 Leu Phe Gly Val ValLeu Leu Thr Leu Val Leu Ser Gly Leu Asn Ala 85 90 95 Thr Leu Ile Gln GlyAla Pro Glu Asp Ser Trp Arg Arg Arg Ile Pro 100 105 110 Ser Ile Phe LeuAsp Val Ala Arg Phe Ala Leu Ile Ala Val Gly Ile 115 120 125 Thr Val IleMet Ala Tyr Val Trp Gly Ala Asn Val Gly Gly Leu Phe 130 135 140 Thr AlaLeu Gly Val Thr Ser Ile Val Leu Gly Leu Ala Leu Gln Asn 145 150 155 160Ser Val Gly Gln Ile Ile Ser Gly Leu Leu Leu Leu Phe Glu Gln Pro 165 170175 Phe Arg Leu Gly Asp Trp Ile Thr Val Pro Thr Ala Ala Gly Arg Pro 180185 190 Ser Ala His Gly Arg Val Val Glu Val Asn Trp Arg Ala Thr His Ile195 200 205 Asp Thr Gly Gly Asn Leu Leu Val Met Pro Asn Ala Glu Leu AlaGly 210 215 220 Ala Ser Phe Thr Asn Tyr Ser Arg Pro Val Gly Glu His ArgLeu Thr 225 230 235 240 Val Val Thr Thr Phe Asn Ala Ala Asp Thr Pro AspAsp Val Cys Glu 245 250 255 Met Leu Ser Ser Val Ala Ala Ser Leu Pro GluLeu Arg Thr Asp Gly 260 265 270 Gln Ile Ala Thr Leu Tyr Leu Gly Ala AlaGlu Tyr Glu Lys Ser Ile 275 280 285 Pro Leu His Thr Pro Ala Val Asp AspSer Val Arg Ser Thr Tyr Leu 290 295 300 Arg Trp Val Trp Tyr Ala Ala ArgArg Gln Glu Leu Arg Xaa Asn Gly 305 310 315 320 Val Ala Asp Xaa Phe AspThr Pro Glu Arg Ile Ala Ser Ala Met Arg 325 330 335 Ala Val Ala Ser ThrLeu Arg Leu Ala Asp Asp Glu Gln Gln Glu Ile 340 345 350 Ala Asp Val ValArg Leu Val Arg Tyr Gly Asn Gly Glu Arg Leu Gln 355 360 365 Gln Pro GlyGln Val Pro Thr Gly Met Arg Phe Ile Val Asp Gly Arg 370 375 380 Val SerLeu Ser Val Ile Asp Gln Asp Gly Asp Val Ile Pro Ala Arg 385 390 395 400Val Leu Glu Arg Gly Asp Phe Leu Gly Gln Thr Thr Leu Thr Arg Glu 405 410415 Pro Val Leu Ala Thr Ala His Ala Leu Glu Glu Val Thr Val Leu Glu 420425 430 Met Ala Arg Asp Glu Ile Glu Arg Leu Val His Arg Lys Pro Ile Leu435 440 445 Leu His Val Ile Gly Ala Val 450 455 <210> SEQ ID NO 122<211> LENGTH: 898 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 122 atgacaattc tgccctggaa tgcgcgaacg tctgaacacccgacgcgaaa aagacgcggg 60 cgctaccacc tcctgtcgcg gatgagcatc cagtccaagttgctgctgat gctgcttctg 120 accagcattc tctcggctgc ggtggtcggt ttcatcggctatcagtccgg acggtcctcg 180 ctgcgcgcat cggtgttcga ccgcctcacc gacatccgcgagtcgcagtc gcgcgggttg 240 gagaatcagt tcgcggacct gaagaactcg atggtgatttactcgcgcgg cagcactgcc 300 acggaggcga tcggcgcgtt cagcgacggt ttccgtcagctcggcgatgc gacgatcaat 360 accgggcagg cggcgtcatt gcgccgttac tacgaccggacgttcgccaa caccaccctc 420 gacgacagcg gaaaccgcgt cgacgtccgc gcgctcatcccgaaatccaa cccccagcgc 480 tatctgcagg cgctctatac cccgccgttt cagaactgggagaaggcgat cgcgttcgac 540 gacgcgcgcg acggcagcgc ctggtcggcc gccaatgccagattcaacga gttcttccgc 600 gagatcgtgc accgcttcaa cttcgaggat ctgatgctgctcgacctcga gggcaacgtg 660 gtgtactccg cctacaaggg gccggatctc gggacaaacatcgtcaacgg cccctatcgc 720 aaccgggaac tgtcggaagc ctacgagaag gcggtcgcgtcgaactcgat cgactatgtc 780 ggtgtcaccg acttcgggtg gtacctgcct gccgaggaaccgaccgcctg gttcctgtcc 840 ccggtcgggt tgaaggaccg agtcgacggt gtgatggcggtccagttccc cggaattc 898 <210> SEQ ID NO 123 <211> LENGTH: 1259 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 123cgcaattgat gacggcgcgg ggacagtggc gtgacaccgg gatgggagac accggtgaga 60ccatcctggt cggaccggac aatctgatgc gctcggactc ccggctgttc cgcgagaacc 120gggagaagtt cctggccgac gtcgtcgagg ggggaacccc gccggaggtc gccgacgaat 180cggttgaccg ccgcggcacc acgctggtgc agccggtgac cacccgctcc gtcgaggagg 240cccaacgcgg caacaccggg acgacgatcg aggacgacta tctcggccac gaggcgttac 300aggcgtactc accggtggac ctgccgggac tgcactgggt gatcgtggcc aagatcgaca 360ccgacgaggc gttcgccccg gtggcgcagt tcaccaggac cctggtgctg tcgacggtga 420tcatcatctt cggcgtgtcg ctggcggcca tgctgctggc gcggttgttc gtccgtccga 480tccggcggtt gcaggccggc gcccagcaga tcagcggcgg tgactaccgc ctcgctctgc 540cggtgttgtc tcgtgacgaa ttcggcgatc tgacaacagc tttcaacgac atgagtcgca 600atctgtcgat caaggacgag ctgctcggcg aggagcgcgc cgagaaccaa cggctgatgc 660tgtccctgat gcccgaaccg gtgatgcagc gctacctcga cggggaggag acgatcgccc 720aggaccacaa gaacgtcacg gtgatcttcg ccgacatgat gggcctcgac gagttgtcgc 780gcatgttgac ctccgaggaa ctgatggtgg tggtcaacga cctgacccgc cagttcgacg 840ccgccgccga gagtctcggg gtcgaccacg tgcggacgct gcacgacggg tacctggcca 900gctgcgggtt aggcgtgccg cggctggaca acgtccggcg cacggtcaat ttcgcgatcg 960aaatggaccg catcatcgac cggcacgccg ccgagtccgg gcacgacctg cggctccgcg 1020cgggcatcga caccgggtcg gcggccagcg ggctggtggg gcggtccacg ttggcgtacg 1080acatgtgggg ttcggcggtc gatgtcgcct accaggtgca gcgcggctcc ccccagcccg 1140gcatctacgt cacctcgcgg gtgcacgagg tcatgcagga aactctcgac ttcgtcgccg 1200ccggggaggt cgtcggcgag cgcggcgtcg agacggtctg gcggttgcag ggccacccg 1259<210> SEQ ID NO 124 <211> LENGTH: 299 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 124 Met Thr Ile Leu Pro Trp Asn AlaArg Thr Ser Glu His Pro Thr Arg 1 5 10 15 Lys Arg Arg Gly Arg Tyr HisLeu Leu Ser Arg Met Ser Ile Gln Ser 20 25 30 Lys Leu Leu Leu Met Leu LeuLeu Thr Ser Ile Leu Ser Ala Ala Val 35 40 45 Val Gly Phe Ile Gly Tyr GlnSer Gly Arg Ser Ser Leu Arg Ala Ser 50 55 60 Val Phe Asp Arg Leu Thr AspIle Arg Glu Ser Gln Ser Arg Gly Leu 65 70 75 80 Glu Asn Gln Phe Ala AspLeu Lys Asn Ser Met Val Ile Tyr Ser Arg 85 90 95 Gly Ser Thr Ala Thr GluAla Ile Gly Ala Phe Ser Asp Gly Phe Arg 100 105 110 Gln Leu Gly Asp AlaThr Ile Asn Thr Gly Gln Ala Ala Ser Leu Arg 115 120 125 Arg Tyr Tyr AspArg Thr Phe Ala Asn Thr Thr Leu Asp Asp Ser Gly 130 135 140 Asn Arg ValAsp Val Arg Ala Leu Ile Pro Lys Ser Asn Pro Gln Arg 145 150 155 160 TyrLeu Gln Ala Leu Tyr Thr Pro Pro Phe Gln Asn Trp Glu Lys Ala 165 170 175Ile Ala Phe Asp Asp Ala Arg Asp Gly Ser Ala Trp Ser Ala Ala Asn 180 185190 Ala Arg Phe Asn Glu Phe Phe Arg Glu Ile Val His Arg Phe Asn Phe 195200 205 Glu Asp Leu Met Leu Leu Asp Leu Glu Gly Asn Val Val Tyr Ser Ala210 215 220 Tyr Lys Gly Pro Asp Leu Gly Thr Asn Ile Val Asn Gly Pro TyrArg 225 230 235 240 Asn Arg Glu Leu Ser Glu Ala Tyr Glu Lys Ala Val AlaSer Asn Ser 245 250 255 Ile Asp Tyr Val Gly Val Thr Asp Phe Gly Trp TyrLeu Pro Ala Glu 260 265 270 Glu Pro Thr Ala Trp Phe Leu Ser Pro Val GlyLeu Lys Asp Arg Val 275 280 285 Asp Gly Val Met Ala Val Gln Phe Pro GlyIle 290 295 <210> SEQ ID NO 125 <211> LENGTH: 419 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 125 Gln Leu Met Thr AlaArg Gly Gln Trp Arg Asp Thr Gly Met Gly Asp 1 5 10 15 Thr Gly Glu ThrIle Leu Val Gly Pro Asp Asn Leu Met Arg Ser Asp 20 25 30 Ser Arg Leu PheArg Glu Asn Arg Glu Lys Phe Leu Ala Asp Val Val 35 40 45 Glu Gly Gly ThrPro Pro Glu Val Ala Asp Glu Ser Val Asp Arg Arg 50 55 60 Gly Thr Thr LeuVal Gln Pro Val Thr Thr Arg Ser Val Glu Glu Ala 65 70 75 80 Gln Arg GlyAsn Thr Gly Thr Thr Ile Glu Asp Asp Tyr Leu Gly His 85 90 95 Glu Ala LeuGln Ala Tyr Ser Pro Val Asp Leu Pro Gly Leu His Trp 100 105 110 Val IleVal Ala Lys Ile Asp Thr Asp Glu Ala Phe Ala Pro Val Ala 115 120 125 GlnPhe Thr Arg Thr Leu Val Leu Ser Thr Val Ile Ile Ile Phe Gly 130 135 140Val Ser Leu Ala Ala Met Leu Leu Ala Arg Leu Phe Val Arg Pro Ile 145 150155 160 Arg Arg Leu Gln Ala Gly Ala Gln Gln Ile Ser Gly Gly Asp Tyr Arg165 170 175 Leu Ala Leu Pro Val Leu Ser Arg Asp Glu Phe Gly Asp Leu ThrThr 180 185 190 Ala Phe Asn Asp Met Ser Arg Asn Leu Ser Ile Lys Asp GluLeu Leu 195 200 205 Gly Glu Glu Arg Ala Glu Asn Gln Arg Leu Met Leu SerLeu Met Pro 210 215 220 Glu Pro Val Met Gln Arg Tyr Leu Asp Gly Glu GluThr Ile Ala Gln 225 230 235 240 Asp His Lys Asn Val Thr Val Ile Phe AlaAsp Met Met Gly Leu Asp 245 250 255 Glu Leu Ser Arg Met Leu Thr Ser GluGlu Leu Met Val Val Val Asn 260 265 270 Asp Leu Thr Arg Gln Phe Asp AlaAla Ala Glu Ser Leu Gly Val Asp 275 280 285 His Val Arg Thr Leu His AspGly Tyr Leu Ala Ser Cys Gly Leu Gly 290 295 300 Val Pro Arg Leu Asp AsnVal Arg Arg Thr Val Asn Phe Ala Ile Glu 305 310 315 320 Met Asp Arg IleIle Asp Arg His Ala Ala Glu Ser Gly His Asp Leu 325 330 335 Arg Leu ArgAla Gly Ile Asp Thr Gly Ser Ala Ala Ser Gly Leu Val 340 345 350 Gly ArgSer Thr Leu Ala Tyr Asp Met Trp Gly Ser Ala Val Asp Val 355 360 365 AlaTyr Gln Val Gln Arg Gly Ser Pro Gln Pro Gly Ile Tyr Val Thr 370 375 380Ser Arg Val His Glu Val Met Gln Glu Thr Leu Asp Phe Val Ala Ala 385 390395 400 Gly Glu Val Val Gly Glu Arg Gly Val Glu Thr Val Trp Arg Leu Gln405 410 415 Gly His Pro <210> SEQ ID NO 126 <211> LENGTH: 27 <212> TYPE:DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 126 ccggatccga tgagcagcgtgctgaac 27 <210> SEQ ID NO 127 <211> LENGTH: 26 <212> TYPE: DNA <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Made in a lab <400> SEQUENCE: 127 gcggatccca cggccccgat cacgtg 26 <210>SEQ ID NO 128 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 128 ccggatccaa tgacatttct gccctggaat gcg 33 <210>SEQ ID NO 129 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 129 ccggatccat tcggtggccc tgcaaccgcc ag 32 <210> SEQID NO 130 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Made in a lab <400>SEQUENCE: 130 ccggatccgg agcaaccgtt ccggctc 27 <210> SEQ ID NO 131 <211>LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Made in a lab <400> SEQUENCE: 131ccggatcccg gctatcagtc cggacgg 27 <210> SEQ ID NO 132 <211> LENGTH: 844<212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 132gagcaaccgt tccggctcgg cgactggatc accgtcccca ccgcggcggg ccggccgtcc 60gcccacggcc gcgtggtgga agtcaactgg cgtgcaacac atatcgacac cggcggcaac 120ctgctggtaa tgcccaacgc cgaactcgcc ggcgcgtcgt tcaccaatta cagccggccc 180gtgggagagc accggctgac cgtcgtcacc accttcaacg ccgcggacac ccccgatgat 240gtctgcgaga tgctgtcgtc ggtcgcggcg tcgctgcccg aactgcgcac cgacggacag 300atcgccacgc tctatctcgg tgcggccgaa tacgagaagt cgatcccgtt gcacacaccc 360gcggtggacg actcggtcag gagcacgtac ctgcgatggg tctggtacgc cgcgcgccgg 420caggaacttc gcctaacggc gtcgccgacg attcgacacg ccggaacgga tcgcctcggc 480catgcgggct gtggcgtcca cactgcgctt ggcagacgac gaacagcagg agatcgccga 540cgtggtgcgt ctggtccgtt acggcaacgg ggaacgcctc cagcagccgg gtcaggtacc 600gaccgggatg aggttcatcg tagacggcag ggtgagtctg tccgtgatcg atcaggacgg 660cgacgtgatc ccggcgcggg tgctcgagcg tggcgacttc ctggggcaga ccacgctgac 720gcgggaaccg gtactggcga ccgcgcacgc gctggaggaa gtcaccgtgc tggagatggc 780ccgtgacgag atcgagcgcc tggtgcaccg aaagccgatc ctgctgcacg tgatcggggc 840cgtg 844 <210> SEQ ID NO 133 <211> LENGTH: 742 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 133 ggctatcagt ccggacggtcctcgctgcgc gcatcggtgt tcgaccgcct caccgacatc 60 cgcgagtcgc agtcgcgcgggttggagaat cagttcgcgg acctgaagaa ctcgatggtg 120 atttactcgc gcggcagcactgccacggag gcgatcggcg cgttcagcga cggtttccgt 180 cagctcggcg atgcgacgatcaataccggg caggcggcgt cattgcgccg ttactacgac 240 cggacgttcg ccaacaccaccctcgacgac agcggaaacc gcgtcgacgt ccgcgcgctc 300 atcccgaaat ccaacccccagcgctatctg caggcgctct ataccccgcc gtttcagaac 360 tgggagaagg cgatcgcgttcgacgacgcg cgcgacggca gcgcctggtc ggccgccaat 420 gccagattca acgagttcttccgcgagatc gtgcaccgct tcaacttcga ggatctgatg 480 ctgctcgacc tcgagggcaacgtggtgtac tccgcctaca aggggccgga tctcgggaca 540 aacatcgtca acggcccctatcgcaaccgg gaactgtcgg aagcctacga gaaggcggtc 600 gcgtcgaact cgatcgactatgtcggtgtc accgacttcg ggtggtacct gcctgccgag 660 gaaccgaccg cctggttcctgtccccggtc gggttgaagg accgagtcga cggtgtgatg 720 gcggtccagt tccccggaat tc742 <210> SEQ ID NO 134 <211> LENGTH: 282 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (145)...(145) <221> NAME/KEY: UNSURE <222> LOCATION:(151)...(151) <400> SEQUENCE: 134 Glu Gln Pro Phe Arg Leu Gly Asp TrpIle Thr Val Pro Thr Ala Ala 1 5 10 15 Gly Arg Pro Ser Ala His Gly ArgVal Val Glu Val Asn Trp Arg Ala 20 25 30 Thr His Ile Asp Thr Gly Gly AsnLeu Leu Val Met Pro Asn Ala Glu 35 40 45 Leu Ala Gly Ala Ser Phe Thr AsnTyr Ser Arg Pro Val Gly Glu His 50 55 60 Arg Leu Thr Val Val Thr Thr PheAsn Ala Ala Asp Thr Pro Asp Asp 65 70 75 80 Val Cys Glu Met Leu Ser SerVal Ala Ala Ser Leu Pro Glu Leu Arg 85 90 95 Thr Asp Gly Gln Ile Ala ThrLeu Tyr Leu Gly Ala Ala Glu Tyr Glu 100 105 110 Lys Ser Ile Pro Leu HisThr Pro Ala Val Asp Asp Ser Val Arg Ser 115 120 125 Thr Tyr Leu Arg TrpVal Trp Tyr Ala Ala Arg Arg Gln Glu Leu Arg 130 135 140 Xaa Asn Gly ValAla Asp Xaa Phe Asp Thr Pro Glu Arg Ile Ala Ser 145 150 155 160 Ala MetArg Ala Val Ala Ser Thr Leu Arg Leu Ala Asp Asp Glu Gln 165 170 175 GlnGlu Ile Ala Asp Val Val Arg Leu Val Arg Tyr Gly Asn Gly Glu 180 185 190Arg Leu Gln Gln Pro Gly Gln Val Pro Thr Gly Met Arg Phe Ile Val 195 200205 Asp Gly Arg Val Ser Leu Ser Val Ile Asp Gln Asp Gly Asp Val Ile 210215 220 Pro Ala Arg Val Leu Glu Arg Gly Asp Phe Leu Gly Gln Thr Thr Leu225 230 235 240 Thr Arg Glu Pro Val Leu Ala Thr Ala His Ala Leu Glu GluVal Thr 245 250 255 Val Leu Glu Met Ala Arg Asp Glu Ile Glu Arg Leu ValHis Arg Lys 260 265 270 Pro Ile Leu Leu His Val Ile Gly Ala Val 275 280<210> SEQ ID NO 135 <211> LENGTH: 247 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 135 Gly Tyr Gln Ser Gly Arg Ser SerLeu Arg Ala Ser Val Phe Asp Arg 1 5 10 15 Leu Thr Asp Ile Arg Glu SerGln Ser Arg Gly Leu Glu Asn Gln Phe 20 25 30 Ala Asp Leu Lys Asn Ser MetVal Ile Tyr Ser Arg Gly Ser Thr Ala 35 40 45 Thr Glu Ala Ile Gly Ala PheSer Asp Gly Phe Arg Gln Leu Gly Asp 50 55 60 Ala Thr Ile Asn Thr Gly GlnAla Ala Ser Leu Arg Arg Tyr Tyr Asp 65 70 75 80 Arg Thr Phe Ala Asn ThrThr Leu Asp Asp Ser Gly Asn Arg Val Asp 85 90 95 Val Arg Ala Leu Ile ProLys Ser Asn Pro Gln Arg Tyr Leu Gln Ala 100 105 110 Leu Tyr Thr Pro ProPhe Gln Asn Trp Glu Lys Ala Ile Ala Phe Asp 115 120 125 Asp Ala Arg AspGly Ser Ala Trp Ser Ala Ala Asn Ala Arg Phe Asn 130 135 140 Glu Phe PheArg Glu Ile Val His Arg Phe Asn Phe Glu Asp Leu Met 145 150 155 160 LeuLeu Asp Leu Glu Gly Asn Val Val Tyr Ser Ala Tyr Lys Gly Pro 165 170 175Asp Leu Gly Thr Asn Ile Val Asn Gly Pro Tyr Arg Asn Arg Glu Leu 180 185190 Ser Glu Ala Tyr Glu Lys Ala Val Ala Ser Asn Ser Ile Asp Tyr Val 195200 205 Gly Val Thr Asp Phe Gly Trp Tyr Leu Pro Ala Glu Glu Pro Thr Ala210 215 220 Trp Phe Leu Ser Pro Val Gly Leu Lys Asp Arg Val Asp Gly ValMet 225 230 235 240 Ala Val Gln Phe Pro Gly Ile 245 <210> SEQ ID NO 136<211> LENGTH: 45 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (18)...(18) <400>SEQUENCE: 136 atgagcgaaa tcgcccgncc ctggcgggtt ctggcatgtg gcatc 45 <210>SEQ ID NO 137 <211> LENGTH: 340 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: unsure <222>LOCATION: (273)...(273) <221> NAME/KEY: unsure <222> LOCATION:(286)...(286) <400> SEQUENCE: 137 gccaccggcg gcgccgccgc ggtgcccgccggggtgagcg ccccggcggt cgcgccggcc 60 cccgcgatgc ccgcccgccc ggtgtccacgatcgcgccgg cgacctcggg cacgctcagc 120 gagtttttcg ccgccaaggg cgtcacgatggagccgcagt ccagccgcga cttccgcgcc 180 ctcaacatcg tgctgccgaa gccgcggggctgggagcaca tcccggaccc gaacgtgccg 240 gacgcgttcg cggtgctggc cgaccgggtcagnggtaaag gtcagnagtc gacaaacgcc 300 cacgtggtgg tcgacaaaca cgtaggcgagttcgacggca 340 <210> SEQ ID NO 138 <211> LENGTH: 235 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (16)...(16) <400> SEQUENCE: 138 ggtgaccaccagcgtngaac aggtcgttgc cgaagccgcg gaggccaccg acgcgattgt 60 caacggcttcaaggtcagcg ttccgggtcc gggtccggcc gcaccgccac ctgcacccgg 120 tgcccccggtgtcccgcccg cccccggcgc cccggcgctg ccgctggccg tcgcaccacc 180 cccggctcccgctgttcccg ccgtggcgcc cgcgccacag ctgctgggac tgcag 235 <210> SEQ ID NO139 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 139 Met Ser Glu Ile Ala Arg Pro Trp Arg Val LeuAla Cys Gly Ile 1 5 10 15 <210> SEQ ID NO 140 <211> LENGTH: 113 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: UNSURE <222> LOCATION: (96)...(96) <400> SEQUENCE: 140 Ala ThrGly Gly Ala Ala Ala Val Pro Ala Gly Val Ser Ala Pro Ala 1 5 10 15 ValAla Pro Ala Pro Ala Met Pro Ala Arg Pro Val Ser Thr Ile Ala 20 25 30 ProAla Thr Ser Gly Thr Leu Ser Glu Phe Phe Ala Ala Lys Gly Val 35 40 45 ThrMet Glu Pro Gln Ser Ser Arg Asp Phe Arg Ala Leu Asn Ile Val 50 55 60 LeuPro Lys Pro Arg Gly Trp Glu His Ile Pro Asp Pro Asn Val Pro 65 70 75 80Asp Ala Phe Ala Val Leu Ala Asp Arg Val Gly Gly Lys Gly Gln Xaa 85 90 95Ser Thr Asn Ala His Val Val Val Asp Lys His Val Gly Glu Phe Asp 100 105110 Gly <210> SEQ ID NO 141 <211> LENGTH: 73 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 141 Val Thr Thr Ser ValGlu Gln Val Val Ala Ala Ala Asp Ala Thr Glu 1 5 10 15 Ala Ile Val AsnGly Phe Lys Val Ser Val Pro Gly Pro Gly Pro Ala 20 25 30 Ala Pro Pro ProAla Pro Gly Ala Pro Gly Val Pro Pro Ala Pro Gly 35 40 45 Ala Pro Ala LeuPro Leu Ala Val Ala Pro Pro Pro Ala Pro Ala Val 50 55 60 Pro Ala Val AlaPro Ala Pro Gln Leu 65 70 <210> SEQ ID NO 142 <211> LENGTH: 273 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 142gcgacctacg tgcagggggg tctcggccgc atcgaggccc gggtggccga cagcggatac 60agcaacgccg cggccaaggg ctacttcccg ctgagcttca ccgtcgccgg catcgaccag 120aacggtccga tcgtgaccgc caacgtcacc gcggcggccc cgacgggcgc cgtggccacc 180cagccgctga cgttcatcgc cgggccgagc ccgaccggat ggcagctgtc caagcagtcc 240gcactggccc tgatgtccgc ggtcatcgcc gca 273 <210> SEQ ID NO 143 <211>LENGTH: 91 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 143 Ala Thr Tyr Val Gln Gly Gly Leu Gly Arg Ile Glu Ala ArgVal Ala 1 5 10 15 Asp Ser Gly Tyr Ser Asn Ala Ala Ala Lys Gly Tyr PhePro Leu Ser 20 25 30 Phe Thr Val Ala Gly Ile Asp Gln Asn Gly Pro Ile ValThr Ala Asn 35 40 45 Val Thr Ala Ala Ala Pro Thr Gly Ala Val Ala Thr GlnPro Leu Thr 50 55 60 Phe Ile Ala Gly Pro Ser Pro Thr Gly Trp Gln Leu SerLys Gln Ser 65 70 75 80 Ala Leu Ala Leu Met Ser Ala Val Ile Ala Ala 8590 <210> SEQ ID NO 144 <211> LENGTH: 554 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 144 gatgtcacgc ccggagaatgtaacgttcga ccggagaacg ccgtcggcac aacgagttac 60 gtttgagcac ttcagatctcggttaccttg gatttcaggc gggggaagca gtaaccgatc 120 caagattcga aggacccaaacaacatgaaa ttcactggaa tgaccgtgcg cgcaagccgc 180 gcgccctggc cggcgtcggggcggcatgtc tgttcggcgg cgtggccgcg gcaaccgtgg 240 cggcacagat ggcgggcgcccagccggccg agtgcaacgc cagctcactc accggcaccg 300 tcagctcggt gaccggtcaggcgcgtcagt acctagacac ccacccgggc gccaaccagg 360 ccgtcaccgc ggcgatgaaccagccgcggc ccgaggccga ggcgaacctg cggggctact 420 tcaccgccaa cccggcggagtactacgacc tgcggggcat cctcgccccg atcggtgacg 480 cgcagcgcaa ctgcaacatcaccgtgctgc cggtagagct gcagacggcc tacgacacgt 540 tcatggccgg ctga 554<210> SEQ ID NO 145 <211> LENGTH: 136 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 145 Met Lys Phe Thr Gly Met Thr ValArg Ala Ser Arg Arg Ala Leu Ala 1 5 10 15 Gly Val Gly Ala Ala Cys LeuPhe Gly Gly Val Ala Ala Ala Thr Val 20 25 30 Ala Ala Gln Met Ala Gly AlaGln Pro Ala Glu Cys Asn Ala Ser Ser 35 40 45 Leu Thr Gly Thr Val Ser SerVal Thr Gly Gln Ala Arg Gln Tyr Leu 50 55 60 Asp Thr His Pro Gly Ala AsnGln Ala Val Thr Ala Ala Met Asn Gln 65 70 75 80 Pro Arg Pro Glu Ala GluAla Asn Leu Arg Gly Tyr Phe Thr Ala Asn 85 90 95 Pro Ala Glu Tyr Tyr AspLeu Arg Gly Ile Leu Ala Pro Ile Gly Asp 100 105 110 Ala Gln Arg Asn CysAsn Ile Thr Val Leu Pro Val Glu Leu Gln Thr 115 120 125 Ala Tyr Asp ThrPhe Met Ala Gly 130 135 <210> SEQ ID NO 146 <211> LENGTH: 808 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221>NAME/KEY: unsure <222> LOCATION: (15)...(15) <400> SEQUENCE: 146ccaagtgtga cgcgngtgtg acggtagacg ttccgaccaa tccaacgacg ccgcagctgg 60gaatcacccg tgtgccaatt cagtgcgggc aacggtgtcc gtccacgaag ggattcagga 120aatgatgaca actcgccgga agtcagccgc agtggcggga atcgctgcgg tggccatcct 180cggtgcggcc gcatgttcga gtgaggacgg tgggagcacg gcctcgtcgg ccagcagcac 240ggcctcctcc gcgatggagt ccgcgaccga cgagatgacc acgtcgtcgg cggccccttc 300ggccgaccct gcggccaacc tgatcggctc cggctgcgcg gcctacgccg agcaggtccc 360cgaaggtccc gggtcggtgg ccgggatggc agccgatccg gtgacggtgg cggcgtcgaa 420caacccgatg ctgcagacgc tgtcccaggc gctgtccggc cagctcaatc cgcaggtcaa 480tctcgtcgac accctcgacg gcggtgagtt caccgtgttc gcgccgaccg acgacgcgtt 540cgccaagatc gatccggcca cgctggagac cctcaagacg gactccgaca tgctgaccaa 600catcctgacc taccacgtcg tgcccggcca ggccgcgccc gatcaggtgg tcggcgagca 660tgtgacggtg gagggggcgc cggtcacggt gtccgggatg gccgaccagc tcaaggtcaa 720cgacgcgtcg gtggtgtgcg gtggggtgca gaccgccaac gcgacggtgt atctgatcga 780caccgtgctg atgccgccgg cagcgtag 808 <210> SEQ ID NO 147 <211> LENGTH: 228<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 147Met Met Thr Thr Arg Arg Lys Ser Ala Ala Val Ala Gly Ile Ala Ala 1 5 1015 Val Ala Ile Leu Gly Ala Ala Ala Cys Ser Ser Glu Asp Gly Gly Ser 20 2530 Thr Ala Ser Ser Ala Ser Ser Thr Ala Ser Ser Ala Met Glu Ser Ala 35 4045 Thr Asp Glu Met Thr Thr Ser Ser Ala Ala Pro Ser Ala Asp Pro Ala 50 5560 Ala Asn Leu Ile Gly Ser Gly Cys Ala Ala Tyr Ala Glu Gln Val Pro 65 7075 80 Glu Gly Pro Gly Ser Val Ala Gly Met Ala Ala Asp Pro Val Thr Val 8590 95 Ala Ala Ser Asn Asn Pro Met Leu Gln Thr Leu Ser Gln Ala Leu Ser100 105 110 Gly Gln Leu Asn Pro Gln Val Asn Leu Val Asp Thr Leu Asp GlyGly 115 120 125 Glu Phe Thr Val Phe Ala Pro Thr Asp Asp Ala Phe Ala LysIle Asp 130 135 140 Pro Ala Thr Leu Glu Thr Leu Lys Thr Asp Ser Asp MetLeu Thr Asn 145 150 155 160 Ile Leu Thr Tyr His Val Val Pro Gly Gln AlaAla Pro Asp Gln Val 165 170 175 Val Gly Glu His Val Thr Val Glu Gly AlaPro Val Thr Val Ser Gly 180 185 190 Met Ala Asp Gln Leu Lys Val Asn AspAla Ser Val Val Cys Gly Gly 195 200 205 Val Gln Thr Ala Asn Ala Thr ValTyr Leu Ile Asp Thr Val Leu Met 210 215 220 Pro Pro Ala Ala 225 <210>SEQ ID NO 148 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <221> NAME/KEY: unsure <222> LOCATION: (12)...(12) <221> NAME/KEY:unsure <222> LOCATION: (17)...(17) <400> SEQUENCE: 148 gcsccsgtsggnccggntgy gc 22 <210> SEQ ID NO 149 <211> LENGTH: 21 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <221> NAME/KEY: unsure <222> LOCATION:(10)...(10) <221> NAME/KEY: unsure <222> LOCATION: (13)...(13) <221>NAME/KEY: unsure <222> LOCATION: (16)...(16) <221> NAME/KEY: unsure<222> LOCATION: (20)...(20) <400> SEQUENCE: 149 rtasgcsgcn gtngcnacng g21 <210> SEQ ID NO 150 <211> LENGTH: 102 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 150 gcccccgtcg gccccggctg tgcggcctac gtgcaacaggtgccggacgg gccgggatcg 60 gtgcagggca tggcgagctc gcccgtagcg accgccgcgt at102 <210> SEQ ID NO 151 <211> LENGTH: 683 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 151 gcccgccaac taaaaccgccgatcatccac tgcaggaagg aatctcacga tcatgaacat 60 cagcatgaaa actcttgccggagcgggttt cgcgatgacc gccgccgtcg gtctgtcgct 120 gggtaccgca ggcagcgccgcagccgcgcc ggtcggaccg gggtgtgcgg cctacgtgca 180 acaggtgccg gacgggccgggatcggtgca gggcatggcg agctcgccgg tggccaccgc 240 ggcggccgac aacccgctgctcaccacgct ctcgcaggcg atctcgggtc agctcaaccc 300 gaacgtcaat ctcgtcgacacgttcaacgg cggccagttc accgtgttcg cgccgaccaa 360 tgacgccttc gccaagatcgatccggccac gctggagacc ctcaagaccg attccgacct 420 gctgaccaag atcctcacctaccacgtcgt gcccggccag gccgcgcccg atcaggtggt 480 cggcgagcat gtgacggtggagggggcgcc ggtcacggtg tccgggatgg ccgaccagct 540 caaggtcaac gacgcgtcggtggtgtgcgg tggggtgcag accgccaacg cgacggtgta 600 tctgatcgac accgtgctgatgccgccggc agcgtagccg ggcggcacca cagaagaggg 660 tcccccgcac ccggcctcccccg 683 <210> SEQ ID NO 152 <211> LENGTH: 231 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 152 Asp Thr Val Leu MetPro Pro Ala Asn Asn Arg Arg Ser Ser Thr Ala 1 5 10 15 Gly Arg Asn LeuThr Ile Met Asn Ile Ser Met Lys Thr Leu Ala Gly 20 25 30 Ala Gly Phe AlaMet Thr Ala Ala Val Gly Leu Ser Leu Gly Thr Ala 35 40 45 Gly Ser Ala AlaAla Ala Pro Val Gly Pro Gly Cys Ala Ala Tyr Val 50 55 60 Gln Gln Val ProAsp Gly Pro Gly Ser Val Gln Gly Met Ala Ser Ser 65 70 75 80 Pro Val AlaThr Ala Ala Ala Asp Asn Pro Leu Leu Thr Thr Leu Ser 85 90 95 Gln Ala IleSer Gly Gln Leu Asn Pro Asn Val Asn Leu Val Asp Thr 100 105 110 Phe AsnGly Gly Gln Phe Thr Val Phe Ala Pro Thr Asn Asp Ala Phe 115 120 125 AlaLys Ile Asp Pro Ala Thr Leu Glu Thr Leu Lys Thr Asp Ser Asp 130 135 140Leu Leu Thr Lys Ile Leu Thr Tyr His Val Val Pro Gly Gln Ala Ala 145 150155 160 Pro Asp Gln Val Val Gly Glu His Val Thr Val Glu Gly Ala Pro Val165 170 175 Thr Val Ser Gly Met Ala Asp Gln Leu Lys Val Asn Asp Ala SerVal 180 185 190 Val Cys Gly Gly Val Gln Thr Ala Asn Ala Thr Val Tyr LeuIle Asp 195 200 205 Thr Val Leu Met Pro Pro Ala Ala Pro Gly Gly Thr ThrGlu Glu Gly 210 215 220 Pro Pro His Pro Ala Ser Pro 225 230 <210> SEQ IDNO 153 <211> LENGTH: 1125 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION:(358)...(358) <400> SEQUENCE: 153 atgcaggtgc ggcgtgttct gggcagtgtcggtgcagcag tcgcggtttc ggccgcgtta 60 tggcagacgg gggtttcgat accgaccgcctcagcggatc cgtgtccgga catcgaggtg 120 atcttcgcgc gcgggaccgg tgcggaacccggcctcgggt gggtcggtga tgcgttcgtc 180 aacgcgctgc ggcccaaggt cggtgagcagtcggtgggca cctacgcggt gaactacccg 240 gcaggattcg gacttcgaca aatcggcgcccatgggcgcg gccgacgcat cggggcgggt 300 gcagtggatg gccgacaact gcccggacaccaagcttgtc ctgggcggca tgtcgcangg 360 cgccggcgtc atcgacctga tcaccgtcgatccgcgaccg ctgggccggt tcacccccac 420 cccgatgccg ccccgcgtcg ccgaccacgtggccgccgtt gtggtcttcg gaaatccgtt 480 gcgcgacatc cgtggtggcg gtccgctgccgcagatgagc ggcacctacg ggccgaagtc 540 gatcgatctg tgtgcgctcg acgatccgttctgctcgccc ggcttcaacc tgccggccca 600 cttcgcctac gccgacaacg gcatggtggaggaagccgcg aacttcgccc gcctggaacc 660 gggccagagc gtcgagctgc ccgaggcgccctacctgcac ctgttcgtcc cgcggggcga 720 ggtaacgctg gaggacgccg gaccgctgcgcgaaggcgac gcagtgcgtt tcaccgcatc 780 gggcggccag cgggtgaccg ccaccgcgcccgcggagatc ctcgtctggg agatgcatgc 840 gggactcggt gcggcataag cgaataggagtcctgctggc cggcgcagca ctgctcgccg 900 gatgcacatc cgaacctgga cccgggccgtcggcggcacc ggccccgacg agcacaaccg 960 agagcgcacc cggtcccgga ctcgtcccggtgaccgtcgc ggtcgacgaa cctctggccg 1020 acgcgccgtt cgaccagccc cgggaggccctggtgccgca gggttggacg ctgtcggtgt 1080 gggcgcggac cgcccggccg cggctggccgcgtgggcccc ggacg 1125 <210> SEQ ID NO 154 <211> LENGTH: 748 <212> TYPE:PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (119)...(119) <400> SEQUENCE: 154 Met Gln Val ArgArg Val Leu Gly Ser Val Gly Ala Ala Val Ala Val 1 5 10 15 Ser Ala AlaLeu Trp Gln Thr Gly Val Ser Ile Pro Thr Ala Ser Ala 20 25 30 Asp Pro CysPro Asp Ile Glu Val Ile Phe Ala Arg Gly Thr Gly Ala 35 40 45 Glu Pro GlyLeu Gly Trp Val Gly Asp Ala Phe Val Asn Ala Leu Arg 50 55 60 Pro Lys ValGly Glu Gln Ser Val Gly Thr Tyr Ala Val Asn Tyr Pro 65 70 75 80 Ala GlyPhe Asp Phe Asp Lys Ser Ala Pro Met Gly Ala Ala Asp Ala 85 90 95 Ser GlyArg Val Gln Trp Met Ala Asp Asn Cys Pro Asp Thr Lys Leu 100 105 110 ValLeu Gly Gly Met Ser Xaa Gly Ala Gly Val Ile Asp Leu Ile Thr 115 120 125Val Asp Pro Arg Pro Leu Gly Arg Phe Thr Pro Thr Pro Met Pro Pro 130 135140 Arg Val Ala Asp His Val Ala Ala Val Val Val Phe Gly Asn Pro Leu 145150 155 160 Arg Asp Ile Arg Gly Gly Gly Pro Arg Leu Glu Pro Arg Gly LeuAsn 165 170 175 Met Glu Thr Ser Glu Arg Gly Leu Tyr Thr His Arg Thr TyrArg Gly 180 185 190 Leu Tyr Pro Arg Leu Tyr Ser Ser Glu Arg Ile Leu GluAla Ser Pro 195 200 205 Leu Glu Cys Tyr Ser Ala Leu Ala Leu Glu Ala SerPro Ala Ser Pro 210 215 220 Pro Arg Pro His Glu Cys Tyr Ser Ser Glu ArgPro Arg Gly Leu Tyr 225 230 235 240 Pro His Glu Ala Ser Asn Leu Glu ProArg Ala Leu Ala His Ile Ser 245 250 255 Pro His Glu Ala Leu Ala Thr TyrArg Ala Leu Ala Ala Ser Pro Ala 260 265 270 Ser Asn Gly Leu Tyr Met GluThr Val Ala Leu Gly Leu Gly Leu Ala 275 280 285 Leu Ala Ala Leu Ala AlaSer Asn Pro His Glu Ala Leu Ala Ala Arg 290 295 300 Gly Leu Glu Gly LeuPro Arg Gly Leu Tyr Gly Leu Asn Ser Glu Arg 305 310 315 320 Val Ala LeuGly Leu Leu Glu Pro Arg Gly Leu Ala Leu Ala Pro Arg 325 330 335 Thr TyrArg Leu Glu His Ile Ser Leu Glu Pro His Glu Val Ala Leu 340 345 350 ProArg Ala Arg Gly Gly Leu Tyr Gly Leu Val Ala Leu Thr His Arg 355 360 365Leu Glu Gly Leu Ala Ser Pro Ala Leu Ala Gly Leu Tyr Pro Arg Leu 370 375380 Glu Ala Arg Gly Gly Leu Gly Leu Tyr Ala Ser Pro Ala Leu Ala Val 385390 395 400 Ala Leu Ala Arg Gly Pro His Glu Thr His Arg Ala Leu Ala SerGlu 405 410 415 Arg Gly Leu Tyr Gly Leu Tyr Gly Leu Asn Ala Arg Gly ValAla Leu 420 425 430 Thr His Arg Ala Leu Ala Thr His Arg Ala Leu Ala ProArg Ala Leu 435 440 445 Ala Gly Leu Ile Leu Glu Leu Glu Val Ala Leu ThrArg Pro Gly Leu 450 455 460 Met Glu Thr His Ile Ser Ala Leu Ala Gly LeuTyr Leu Glu Gly Leu 465 470 475 480 Tyr Ala Leu Ala Ala Leu Ala Ala LeuAla Ala Ser Asn Ala Arg Gly 485 490 495 Ser Glu Arg Pro Arg Ala Leu AlaGly Leu Tyr Ala Arg Gly Ala Arg 500 505 510 Gly Ser Glu Arg Thr His ArgAla Leu Ala Ala Arg Gly Ala Arg Gly 515 520 525 Met Glu Thr His Ile SerIle Leu Glu Ala Arg Gly Thr His Arg Thr 530 535 540 Arg Pro Thr His ArgAla Arg Gly Ala Leu Ala Val Ala Leu Gly Leu 545 550 555 560 Tyr Gly LeuTyr Thr His Arg Gly Leu Tyr Pro Arg Ala Ser Pro Gly 565 570 575 Leu HisIle Ser Ala Ser Asn Ala Arg Gly Gly Leu Ala Arg Gly Thr 580 585 590 HisArg Ala Arg Gly Ser Glu Arg Ala Arg Gly Thr His Arg Ala Arg 595 600 605Gly Pro Arg Gly Leu Tyr Ala Ser Pro Ala Arg Gly Ala Arg Gly Gly 610 615620 Leu Tyr Ala Arg Gly Ala Arg Gly Thr His Arg Ser Glu Arg Gly Leu 625630 635 640 Tyr Ala Arg Gly Ala Arg Gly Ala Leu Ala Val Ala Leu Ala ArgGly 645 650 655 Pro Arg Ala Leu Ala Pro Arg Gly Leu Tyr Gly Leu Tyr ProArg Gly 660 665 670 Leu Tyr Ala Leu Ala Ala Leu Ala Gly Leu Tyr Leu GluAla Ser Pro 675 680 685 Ala Leu Ala Val Ala Leu Gly Leu Tyr Val Ala LeuGly Leu Tyr Ala 690 695 700 Leu Ala Ala Ser Pro Ala Arg Gly Pro Arg AlaLeu Ala Ala Leu Ala 705 710 715 720 Ala Leu Ala Gly Leu Tyr Ala Arg GlyVal Ala Leu Gly Leu Tyr Pro 725 730 735 Arg Gly Leu Tyr Ala Arg Gly ProArg Gly Leu Tyr 740 745 <210> SEQ ID NO 155 <211> LENGTH: 666 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 155atgaaggcaa atcattcggg atgctacaaa tccgccggcc cgatatggtc gcatccatcg 60ccgctttgtt cgcccgcact ggcaccatct catgcaggtc tggacaatga gctgagcctg 120ggcatccacg gccagggccc ggaacgactg accattcagc agtgggacac cttcctcaac 180ggcgtcttcc cgttggaccg caaccggttg acccgggagt ggttccactc gggcaaggcg 240acctacgtcg tggccggtga aggtgccgac gagttcgagg gcacgctgga gctgggctac 300caggtgggct ttccgtggtc gctgggcgtg ggcatcaact tcagctacac caccccgaac 360atcacgtacg acggttacgg cctcaacttc gccgacccgc tgctgggctt cggtgattcc 420atcgtgaccc cgccgctgtt cccgggtgtc tcgatcacgg cggacctggg caacggcccc 480ggcatccagg aggtcgcgac cttctccgtg gacgtggccg gccccggtgg ttccgtggtg 540gtgtccaacg cgcacggcac ggtcaccggt gctgccggtg gtgtgctgct gcgtccgttc 600gcccgcctga tctcgtcgac cggcgacagc gtcaccacct acggcgcacc ctggaacatg 660aactga 666 <210> SEQ ID NO 156 <211> LENGTH: 221 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 156 Met Lys Ala Asn HisSer Gly Cys Tyr Lys Ser Ala Gly Pro Ile Trp 1 5 10 15 Ser His Pro SerPro Leu Cys Ser Pro Ala Leu Ala Pro Ser His Ala 20 25 30 Gly Leu Asp AsnGlu Leu Ser Leu Gly Val His Gly Gln Gly Pro Glu 35 40 45 His Leu Thr IleGln Gln Trp Asp Thr Phe Leu Asn Gly Val Phe Pro 50 55 60 Leu Asp Arg AsnArg Leu Thr Arg Glu Trp Phe His Ser Gly Lys Ala 65 70 75 80 Thr Tyr ValVal Ala Gly Glu Gly Ala Asp Glu Phe Glu Gly Thr Leu 85 90 95 Glu Leu GlyTyr His Val Gly Phe Pro Trp Ser Leu Gly Val Gly Ile 100 105 110 Asn PheSer Tyr Thr Thr Pro Asn Ile Thr Tyr Asp Gly Tyr Gly Leu 115 120 125 AsnPhe Ala Asp Pro Leu Leu Gly Phe Gly Asp Ser Ile Val Thr Pro 130 135 140Pro Leu Phe Pro Gly Val Ser Ile Thr Ala Asp Leu Gly Asn Gly Pro 145 150155 160 Gly Ile Gln Glu Val Ala Thr Phe Ser Val Asp Val Ala Gly Pro Gly165 170 175 Gly Ser Val Val Val Ser Asn Ala His Gly Thr Val Thr Gly AlaAla 180 185 190 Gly Gly Val Leu Leu Arg Pro Phe Ala Arg Leu Ile Ser SerThr Gly 195 200 205 Asp Ser Val Thr Thr Tyr Gly Ala Pro Trp Asn Met Asn210 215 220 <210> SEQ ID NO 157 <211> LENGTH: 480 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 157 aacggctggg acatcaacacccctgcgttc gagtggttct acgagtccgg cttgtcgacg 60 atcatgccgg tcggcggacagtccagcttc tacagcgact ggtaccagcc gtctcggggc 120 aacgggcaga actacacctacaagtgggag acgttcctga cccaggagct gccgacgtgg 180 ctggaggcca accgcggagtgtcgcgcacc ggcaacgcgt tcgtcggcct gtcgatggcg 240 ggcagcgcgg cgctgacctacgcgatccat cacccgcagc agttcatcta cgcctcgtcg 300 ctgtcaggct tcctgaacccgtccgagggc tggtggccga tgctgatcgg gctggcgatg 360 aacgacgcag gcggcttcaacgccgagagc atgtggggcc cgtcctcgga cccggcgtgg 420 aagcgcaacg acccgatggtcaacatcaac cagctggtgg ccaacaacac ccggatctgg 480 <210> SEQ ID NO 158<211> LENGTH: 161 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 158 Asn Gly Trp Asp Ile Asn Thr Pro Ala Phe Glu Trp PheTyr Glu Ser 1 5 10 15 Gly Leu Ser Thr Ile Met Pro Val Gly Gly Gln SerSer Phe Tyr Ser 20 25 30 Asp Trp Tyr Gln Pro Ser Arg Gly Asn Gly Gln AsnTyr Thr Tyr Lys 35 40 45 Trp Glu Thr Phe Leu Thr Gln Glu Leu Pro Thr TrpLeu Glu Ala Asn 50 55 60 Arg Gly Val Ser Arg Thr Gly Asn Ala Phe Val GlyLeu Ser Met Ala 65 70 75 80 Gly Ser Ala Ala Leu Thr Tyr Ala Ile His HisPro Gln Gln Phe Ile 85 90 95 Tyr Ala Ser Ser Leu Ser Gly Phe Leu Asn ProSer Glu Gly Trp Trp 100 105 110 Pro Met Leu Ile Gly Leu Ala Met Asn AspAla Gly Gly Phe Asn Ala 115 120 125 Glu Ser Met Trp Gly Pro Ser Ser AspPro Ala Trp Lys Arg Asn Asp 130 135 140 Pro Met Val Asn Ile Asn Gln LeuVal Ala Asn Asn Thr Arg Ile Trp 145 150 155 160 Ile <210> SEQ ID NO 159<211> LENGTH: 1626 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 159 atggccaaga caattgcgta tgacgaagag gcccgccgtggcctcgagcg gggcctcaac 60 gccctcgcag acgccgtaaa ggtgacgttg ggcccgaagggtcgcaacgt cgtgctggag 120 aagaagtggg gcgcccccac gatcaccaac gatggtgtgtccatcgccaa ggagatcgag 180 ctggaggacc cgtacgagaa gatcggcgct gagctggtcaaagaggtcgc caagaagacc 240 gacgacgtcg cgggcgacgg caccaccacc gccaccgtgctcgctcaggc tctggttcgc 300 gaaggcctgc gcaacgtcgc agccggcgcc aacccgctcggcctcaagcg tggcatcgag 360 aaggctgtcg aggctgtcac ccagtcgctg ctgaagtcggccaaggaggt cgagaccaag 420 gagcagattt ctgccaccgc ggcgatttcc gccggcgacacccagatcgg cgagctcatc 480 gccgaggcca tggacaaggt cggcaacgag ggtgtcatcaccgtcgagga gtcgaacacc 540 ttcggcctgc agctcgagct caccgagggt atgcgcttcgacaagggcta catctcgggt 600 tacttcgtga ccgacgccga gcgccaggaa gccgtcctggaggatcccta catcctgctg 660 gtcagctcca aggtgtcgac cgtcaaggat ctgctcccgctgctggagaa ggtcatccag 720 gccggcaagc cgctgctgat catcgccgag gacgtcgagggcgaggccct gtccacgctg 780 gtggtcaaca agatccgcgg caccttcaag tccgtcgccgtcaaggctcc gggcttcggt 840 gaccgccgca aggcgatgct gcaggacatg gccatcctcaccggtggtca ggtcgtcagc 900 gaaagagtcg ggctgtccct ggagaccgcc gacgtctcgctgctgggcca ggcccgcaag 960 gtcgtcgtca ccaaggacga gaccaccatc gtcgagggctcgggcgattc cgatgccatc 1020 gccggccggg tggctcagat ccgcgccgag atcgagaacagcgactccga ctacgaccgc 1080 gagaagctgc aggagcgcct ggccaagctg gccggcggtgttgcggtgat caaggccgga 1140 gctgccaccg aggtggagct caaggagcgc aagcaccgcatcgaggacgc cgtccgcaac 1200 gcgaaggctg ccgtcgaaga gggcatcgtc gccggtggcggcgtggctct gctgcagtcg 1260 gctcctgcgc tggacgacct cggcctgacg ggcgacgaggccaccggtgc caacatcgtc 1320 cgcgtggcgc tgtcggctcc gctcaagcag atcgccttcaacggcggcct ggagcccggc 1380 gtcgttgccg agaaggtgtc caacctgccc gcgggtcacggcctcaacgc cgcgaccggt 1440 gagtacgagg acctgctcaa ggccggcgtc gccgacccggtgaaggtcac ccgctcggcg 1500 ctgcagaacg cggcgtccat cgcggctctg ttcctcaccaccgaggccgt cgtcgccgac 1560 aagccggaga aggcgtccgc acccgcgggc gacccgaccggtggcatggg cggtatggac 1620 ttctaa 1626 <210> SEQ ID NO 160 <211> LENGTH:541 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE:160 Met Ala Lys Thr Ile Ala Tyr Asp Glu Glu Ala Arg Arg Gly Leu Glu 1 510 15 Arg Gly Leu Asn Ala Leu Ala Asp Ala Val Lys Val Thr Leu Gly Pro 2025 30 Lys Gly Arg Asn Val Val Leu Glu Lys Lys Trp Gly Ala Pro Thr Ile 3540 45 Thr Asn Asp Gly Val Ser Ile Ala Lys Glu Ile Glu Leu Glu Asp Pro 5055 60 Tyr Glu Lys Ile Gly Ala Glu Leu Val Lys Glu Val Ala Lys Lys Thr 6570 75 80 Asp Asp Val Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala Gln85 90 95 Ala Leu Val Arg Glu Gly Leu Arg Asn Val Ala Ala Gly Ala Asn Pro100 105 110 Leu Gly Leu Lys Arg Gly Ile Glu Lys Ala Val Glu Ala Val ThrGln 115 120 125 Ser Leu Leu Lys Ser Ala Lys Glu Val Glu Thr Lys Glu GlnIle Ser 130 135 140 Ala Thr Ala Ala Ile Ser Ala Gly Asp Thr Gln Ile GlyGlu Leu Ile 145 150 155 160 Ala Glu Ala Met Asp Lys Val Gly Asn Glu GlyVal Ile Thr Val Glu 165 170 175 Glu Ser Asn Thr Phe Gly Leu Gln Leu GluLeu Thr Glu Gly Met Arg 180 185 190 Phe Asp Lys Gly Tyr Ile Ser Gly TyrPhe Val Thr Asp Ala Glu Arg 195 200 205 Gln Glu Ala Val Leu Glu Asp ProTyr Ile Leu Leu Val Ser Ser Lys 210 215 220 Val Ser Thr Val Lys Asp LeuLeu Pro Leu Leu Glu Lys Val Ile Gln 225 230 235 240 Ala Gly Lys Pro LeuLeu Ile Ile Ala Glu Asp Val Glu Gly Glu Ala 245 250 255 Leu Ser Thr LeuVal Val Asn Lys Ile Arg Gly Thr Phe Lys Ser Val 260 265 270 Ala Val LysAla Pro Gly Phe Gly Asp Arg Arg Lys Ala Met Leu Gln 275 280 285 Asp MetAla Ile Leu Thr Gly Gly Gln Val Val Ser Glu Arg Val Gly 290 295 300 LeuSer Leu Glu Thr Ala Asp Val Ser Leu Leu Gly Gln Ala Arg Lys 305 310 315320 Val Val Val Thr Lys Asp Glu Thr Thr Ile Val Glu Gly Ser Gly Asp 325330 335 Ser Asp Ala Ile Ala Gly Arg Val Ala Gln Ile Arg Ala Glu Ile Glu340 345 350 Asn Ser Asp Ser Asp Tyr Asp Arg Glu Lys Leu Gln Glu Arg LeuAla 355 360 365 Lys Leu Ala Gly Gly Val Ala Val Ile Lys Ala Gly Ala AlaThr Glu 370 375 380 Val Glu Leu Lys Glu Arg Lys His Arg Ile Glu Asp AlaVal Arg Asn 385 390 395 400 Ala Lys Ala Ala Val Glu Glu Gly Ile Val AlaGly Gly Gly Val Ala 405 410 415 Leu Leu Gln Ser Ala Pro Ala Leu Asp AspLeu Gly Leu Thr Gly Asp 420 425 430 Glu Ala Thr Gly Ala Asn Ile Val ArgVal Ala Leu Ser Ala Pro Leu 435 440 445 Lys Gln Ile Ala Phe Asn Gly GlyLeu Glu Pro Gly Val Val Ala Glu 450 455 460 Lys Val Ser Asn Leu Pro AlaGly His Gly Leu Asn Ala Ala Thr Gly 465 470 475 480 Glu Tyr Glu Asp LeuLeu Lys Ala Gly Val Ala Asp Pro Val Lys Val 485 490 495 Thr Arg Ser AlaLeu Gln Asn Ala Ala Ser Ile Ala Ala Leu Phe Leu 500 505 510 Thr Thr GluAla Val Val Ala Asp Lys Pro Glu Lys Ala Ser Ala Pro 515 520 525 Ala GlyAsp Pro Thr Gly Gly Met Gly Gly Met Asp Phe 530 535 540 <210> SEQ ID NO161 <211> LENGTH: 985 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 161 ggatccctac atcctgctgg tcagctccaa ggtgtcgaccgtcaaggatc tgctcccgct 60 gctggagaag gtcatccagg ccggcaagcc gctgctgatcatcgccgagg acgtcgaggg 120 cgaggccctg tccacgctgg tggtcaacaa gatccgcggcaccttcaagt ccgtcgccgt 180 caaggctccg ggcttcggtg accgccgcaa ggcgatgctgcaggacatgg ccatcctcac 240 cggtggtcag gtcgtcagcg aaagagtcgg gctgtccctggagaccgccg acgtctcgct 300 gctgggccag gcccgcaagg tcgtcgtcac caaggacgagaccaccatcg tcgagggctc 360 gggcgattcc gatgccatcg ccggccgggt ggctcagatccgcgccgaga tcgagaacag 420 cgactccgac tacgaccgcg agaagctgca ggagcgcctggccaagctgg ccggcggtgt 480 tgcggtgatc aaggccggag ctgccaccga ggtggagctcaaggagcgca agcaccgcat 540 cgaggacgcc gtccgcaacg cgaaggctgc cgtcgaagagggcatcgtcg ccggtggcgg 600 cgtggctctg ctgcagtcgg ctcctgcgct ggacgacctcggcctgacgg gcgacgaggc 660 caccggtgcc aacatcgtcc gcgtggcgct gtcggctccgctcaagcaga tcgccttcaa 720 cggcggcctg gagcccggcg tcgttgccga gaaggtgtccaacctgcccg cgggtcacgg 780 cctcaacgcc gcgaccggtg agtacgagga cctgctcaaggccggcgtcg ccgacccggt 840 gaaggtcacc cgctcggcgc tgcagaacgc ggcgtccatcgcggctctgt tcctcaccac 900 cgaggccgtc gtcgccgaca agccggagaa ggcgtccgcacccgcgggcg acccgaccgg 960 tggcatgggc ggtatggact tctaa 985 <210> SEQ IDNO 162 <211> LENGTH: 327 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 162 Asp Pro Tyr Ile Leu Leu Val Ser Ser Lys ValSer Thr Val Lys Asp 1 5 10 15 Leu Leu Pro Leu Leu Glu Lys Val Ile GlnAla Gly Lys Pro Leu Leu 20 25 30 Ile Ile Ala Glu Asp Val Glu Gly Glu AlaLeu Ser Thr Leu Val Val 35 40 45 Asn Lys Ile Arg Gly Thr Phe Lys Ser ValAla Val Lys Ala Pro Gly 50 55 60 Phe Gly Asp Arg Arg Lys Ala Met Leu GlnAsp Met Ala Ile Leu Thr 65 70 75 80 Gly Gly Gln Val Val Ser Glu Arg ValGly Leu Ser Leu Glu Thr Ala 85 90 95 Asp Val Ser Leu Leu Gly Gln Ala ArgLys Val Val Val Thr Lys Asp 100 105 110 Glu Thr Thr Ile Val Glu Gly SerGly Asp Ser Asp Ala Ile Ala Gly 115 120 125 Arg Val Ala Gln Ile Arg AlaGlu Ile Glu Asn Ser Asp Ser Asp Tyr 130 135 140 Asp Arg Glu Lys Leu GlnGlu Arg Leu Ala Lys Leu Ala Gly Gly Val 145 150 155 160 Ala Val Ile LysAla Gly Ala Ala Thr Glu Val Glu Leu Lys Glu Arg 165 170 175 Lys His ArgIle Glu Asp Ala Val Arg Asn Ala Lys Ala Ala Val Glu 180 185 190 Glu GlyIle Val Ala Gly Gly Gly Val Ala Leu Leu Gln Ser Ala Pro 195 200 205 AlaLeu Asp Asp Leu Gly Leu Thr Gly Asp Glu Ala Thr Gly Ala Asn 210 215 220Ile Val Arg Val Ala Leu Ser Ala Pro Leu Lys Gln Ile Ala Phe Asn 225 230235 240 Gly Gly Leu Glu Pro Gly Val Val Ala Glu Lys Val Ser Asn Leu Pro245 250 255 Ala Gly His Gly Leu Asn Ala Ala Thr Gly Glu Tyr Glu Asp LeuLeu 260 265 270 Lys Ala Gly Val Ala Asp Pro Val Lys Val Thr Arg Ser AlaLeu Gln 275 280 285 Asn Ala Ala Ser Ile Ala Ala Leu Phe Leu Thr Thr GluAla Val Val 290 295 300 Ala Asp Lys Pro Glu Lys Ala Ser Ala Pro Ala GlyAsp Pro Thr Gly 305 310 315 320 Gly Met Gly Gly Met Asp Phe 325 <210>SEQ ID NO 163 <211> LENGTH: 403 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 163 ggatccgcgg caccggctggtgacgaccaa gtacaacccg gcccgcacct ggacggccga 60 gaactccgtc ggcatcggcggcgcgtacct gtgcatctac gggatggagg gccccggcgg 120 ctatcagttc gtcggccgcaccacccaggt gtggagtcgt taccgccaca cggcgccgtt 180 cgaacccgga agtccctggctgctgcggtt tttcgaccga atttcgtggt atccggtgtc 240 ggccgaggag ctgctggaattgcgagccga catggccgca ggccggggct cggtcgacat 300 caccgacggc gtgttctccctcgccgagca cgaacggttc ctggccgaca acgccgacga 360 catcgccgcg ttccgttcccggcaggcggc cgcgttctcc gcc 403 <210> SEQ ID NO 164 <211> LENGTH: 336<212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 164cggaccgcgt gggcggccgc cggcgagttc gaccgcgccg agaaagccgc gtcgaaggcc 60accgacgccg ataccgggga cctggtgctc tacgacggtg cgagcgggtc gacgctccgt 120tcgcgtcgag cgtgtggaag gtcgacgtcg ccgtcggtga ccgggtggtg gccggacagc 180cgttgctggc gctggaggcg atgaagatgg agaccgtgct gcgcgccccg gccgacgggg 240tggtcaccca gatcctggtc tccgctgggc atctcgtcga tcccggcacc ccactggtcg 300tggtcggcac cggagtgcgc gcatgagcgc cgtcga 336 <210> SEQ ID NO 165 <211>LENGTH: 134 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400>SEQUENCE: 165 Asp Pro Arg His Arg Leu Val Thr Thr Lys Tyr Asn Pro AlaArg Thr 1 5 10 15 Trp Thr Ala Glu Asn Ser Val Gly Ile Gly Gly Ala TyrLeu Cys Ile 20 25 30 Tyr Gly Met Glu Gly Pro Gly Gly Tyr Gln Phe Val GlyArg Thr Thr 35 40 45 Gln Val Trp Ser Arg Tyr Arg His Thr Ala Pro Phe GluPro Gly Ser 50 55 60 Pro Trp Leu Leu Arg Phe Phe Asp Arg Ile Ser Trp TyrPro Val Ser 65 70 75 80 Ala Glu Glu Leu Leu Glu Leu Arg Ala Asp Met AlaAla Gly Arg Gly 85 90 95 Ser Val Asp Ile Thr Asp Gly Val Phe Ser Leu AlaGlu His Glu Arg 100 105 110 Phe Leu Ala Asp Asn Ala Asp Asp Ile Ala AlaPhe Arg Ser Arg Gln 115 120 125 Ala Ala Ala Phe Ser Ala 130 <210> SEQ IDNO 166 <211> LENGTH: 108 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 166 Arg Thr Ala Trp Ala Ala Ala Gly Glu Phe AspArg Ala Glu Lys Ala 1 5 10 15 Ala Ser Lys Ala Thr Asp Ala Asp Thr GlyAsp Leu Val Leu Tyr Asp 20 25 30 Gly Asp Glu Arg Val Asp Ala Pro Phe AlaSer Ser Val Trp Lys Val 35 40 45 Asp Val Ala Val Gly Asp Arg Val Val AlaGly Gln Pro Leu Leu Ala 50 55 60 Leu Glu Ala Met Lys Met Glu Thr Val LeuArg Ala Pro Ala Asp Gly 65 70 75 80 Val Val Thr Gln Ile Leu Val Ser AlaGly His Leu Val Asp Pro Gly 85 90 95 Thr Pro Leu Val Val Val Gly Thr GlyVal Arg Ala 100 105 <210> SEQ ID NO 167 <211> LENGTH: 31 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 167 atagaattcg tccgacagtgggacctcgag c 31 <210> SEQ ID NO 168 <211> LENGTH: 27 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <400> SEQUENCE: 168 atagaattcc caccgcgtcagccgccg 27 <210> SEQ ID NO 169 <211> LENGTH: 1111 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 169 gtccgacagt gggacctcgagcaccacgtc acaggacagc ggccccgcca gcggcgccct 60 gcgcgtctcc aactggccgctctatatggc cgacggtttc atcgcagcgt tccagaccgc 120 ctcgggcatc acggtcgactacaaagaaga cttcaacgac aacgagcagt ggttcgccaa 180 ggtcaaggag ccgttgtcgcgcaagcagga cataggcgcc gacctggtga tccccaccga 240 gttcatggcc gcgcgcgtcaagggcctggg atggctcaat gagatcagcg aagccggcgt 300 gcccaatcgc aagaatctgcgtcaggacct gttggactcg agcatcgacg agggccgcaa 360 gttcaccgcg ccgtacatgaccggcatggt cggtctcgcc tacaacaagg cagccaccgg 420 acgcgatatc cgcaccatcgacgacctctg ggatcccgcg ttcaagggcc gcgtcagtct 480 gttctccgac gtccaggacggcctcggcat gatcatgctc tcgcagggca actcgccgga 540 gaatccgacc accgagtccattcagcaggc ggtcgatctg gtccgcgaac agaacgacag 600 ggggtcagat ccgtcgcttcaccggcaacg actacgccga cgacctggcc gcagaaacat 660 cgccatcgcg caggcgtactccggtgacgt cgtgcagctg caggcggaca accccgatct 720 gcagttcatc gttcccgaatccggcggcga ctggttcgtc gacacgatgg tgatcccgta 780 caccacgcag aaccagaaggccgccgaggc gtggatcgac tacatctacg accgagccaa 840 ctacgccaag ctggtcgcgttcacccagtt cgtgcccgca ctctcggaca tgaccgacga 900 actcgccaag gtcgatcctgcatcggcgga gaacccgctg atcaacccgt cggccgaggt 960 gcaggcgaac ctgaagtcgtgggcggcact gaccgacgag cagacgcagg agttcaacac 1020 tgcgtacgcc gccgtcaccggcggctgacg cggtggtagt gccgatgcga ggggcataaa 1080 tggccctgcg gacgcgaggagcataaatgg c 1111 <210> SEQ ID NO 170 <211> LENGTH: 348 <212> TYPE: PRT<213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 170 Ser Asp Ser GlyThr Ser Ser Thr Thr Ser Gln Asp Ser Gly Pro Ala 1 5 10 15 Ser Gly AlaLeu Arg Val Ser Asn Trp Pro Leu Tyr Met Ala Asp Gly 20 25 30 Phe Ile AlaAla Phe Gln Thr Ala Ser Gly Ile Thr Val Asp Tyr Lys 35 40 45 Glu Asp PheAsn Asp Asn Glu Gln Trp Phe Ala Lys Val Lys Glu Pro 50 55 60 Leu Ser ArgLys Gln Asp Ile Gly Ala Asp Leu Val Ile Pro Thr Glu 65 70 75 80 Phe MetAla Ala Arg Val Lys Gly Leu Gly Trp Leu Asn Glu Ile Ser 85 90 95 Glu AlaGly Val Pro Asn Arg Lys Asn Leu Arg Gln Asp Leu Leu Asp 100 105 110 SerSer Ile Asp Glu Gly Arg Lys Phe Thr Ala Pro Tyr Met Thr Gly 115 120 125Met Val Gly Leu Ala Tyr Asn Lys Ala Ala Thr Gly Arg Asp Ile Arg 130 135140 Thr Ile Asp Asp Leu Trp Asp Pro Ala Phe Lys Gly Arg Val Ser Leu 145150 155 160 Phe Ser Asp Val Gln Asp Gly Leu Gly Met Ile Met Leu Ser GlnGly 165 170 175 Asn Ser Pro Glu Asn Pro Thr Thr Glu Ser Ile Gln Gln AlaVal Asp 180 185 190 Leu Val Arg Glu Gln Asn Asp Arg Gly Gln Ile Arg ArgPhe Thr Gly 195 200 205 Asn Asp Tyr Ala Asp Asp Leu Ala Ala Gly Asn IleAla Ile Ala Gln 210 215 220 Ala Tyr Ser Gly Asp Val Val Gln Leu Gln AlaAsp Asn Pro Asp Leu 225 230 235 240 Gln Phe Ile Val Pro Glu Ser Gly GlyAsp Trp Phe Val Asp Thr Met 245 250 255 Val Ile Pro Tyr Thr Thr Gln AsnGln Lys Ala Ala Glu Ala Trp Ile 260 265 270 Asp Tyr Ile Tyr Asp Arg AlaAsn Tyr Ala Lys Leu Val Ala Phe Thr 275 280 285 Gln Phe Val Pro Ala LeuSer Asp Met Thr Asp Glu Leu Ala Lys Val 290 295 300 Asp Pro Ala Ser AlaGlu Asn Pro Leu Ile Asn Pro Ser Ala Glu Val 305 310 315 320 Gln Ala AsnLeu Lys Ser Trp Ala Ala Leu Thr Asp Glu Gln Thr Gln 325 330 335 Glu PheAsn Thr Ala Tyr Ala Ala Val Thr Gly Gly 340 345 <210> SEQ ID NO 171<211> LENGTH: 1420 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION: (955)...(955)<221> NAME/KEY: unsure <222> LOCATION: (973)...(973) <400> SEQUENCE: 171gatgagcagc gtgctgaact cgacctggtt ggcctgggcc gtcgcggtcg cggtcgggtt 60cccggtgctg ctggtcgtgc tgaccgaggt gcacaacgcg ttgcgtcggc gcggcagcgc 120gctggcccgc ccggtgcaac tcctgcgtac ctacatcctg ccgctgggcg cgttgctgct 180cctgctggta caggcgatgg agatctccga cgacgccacg tcggtacggt tggtcgccac 240cctgttcggc gtcgtgttgt tgacgttggt gctgtccggg ctcaacgcca ccctcatcca 300gggcgcacca gaagacagct ggcgcaggcg gattccgtcg atcttcctcg acgtcgcgcg 360cttcgcgctg atcgcggtcg gtatcaccgt gatcatggcc tatgtctggg gcgcgaacgt 420ggggggcctg ttcaccgcac tgggcgtcac ttccatcgtt cttggcctgg ctctgcagaa 480ttcggtcggt cagatcatct cgggtctgct gctgctgttc gagcaaccgt tccggctcgg 540cgactggatc accgtcccca ccgcggcggg ccggccgtcc gcccacggcc gcgtggtgga 600agtcaactgg cgtgcaacac atatcgacac cggcggcaac ctgctggtaa tgcccaacgc 660cgaactcgcc ggcgcgtcgt tcaccaatta cagccggccc gtgggagagc accggctgac 720cgtcgtcacc accttcaacg ccgcggacac ccccgatgat gtctgcgaga tgctgtcgtc 780ggtcgcggcg tcgctgcccg aactgcgcac cgacggacag atcgccacgc tctatctcgg 840tgcggccgaa tacgagaagt cgatcccgtt gcacacaccc gcggtggacg actcggtcag 900gagcacgtac ctgcgatggg tctggtacgc cgcgcgccgg caggaacttc gcctnaacgg 960cgtcgccgac ganttcgaca cgccggaacg gatcgcctcg gccatgcggg ctgtggcgtc 1020cacactgcgc ttggcagacg acgaacagca ggagatcgcc gacgtggtgc gtctggtccg 1080ttacggcaac ggggaacgcc tccagcagcc gggtcaggta ccgaccggga tgaggttcat 1140cgtagacggc agggtgagtc tgtccgtgat cgatcaggac ggcgacgtga tcccggcgcg 1200ggtgctcgag cgtggcgact tcctggggca gaccacgctg acgcgggaac cggtactggc 1260gaccgcgcac gcgctggagg aagtcaccgt gctggagatg gcccgtgacg agatcgagcg 1320cctggtgcac cgaaagccga tcctgctgca cgtgatcggg gccgtgatcg ccgaccggcg 1380cgcgcacgaa cttcggttga tggcggactc gcaggactga 1420 <210> SEQ ID NO 172<211> LENGTH: 471 <212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae<220> FEATURE: <221> NAME/KEY: UNSURE <222> LOCATION: (318)...(318)<221> NAME/KEY: UNSURE <222> LOCATION: (324)...(324) <400> SEQUENCE: 172Met Ser Ser Val Leu Asn Ser Thr Trp Leu Ala Trp Ala Val Ala Val 1 5 1015 Ala Val Gly Phe Pro Val Leu Leu Val Val Leu Thr Glu Val His Asn 20 2530 Ala Leu Arg Arg Arg Gly Ser Ala Leu Ala Arg Pro Val Gln Leu Leu 35 4045 Arg Thr Tyr Ile Leu Pro Leu Gly Ala Leu Leu Leu Leu Leu Val Gln 50 5560 Ala Met Glu Ile Ser Asp Asp Ala Thr Ser Val Arg Leu Val Ala Thr 65 7075 80 Leu Phe Gly Val Val Leu Leu Thr Leu Val Leu Ser Gly Leu Asn Ala 8590 95 Thr Leu Ile Gln Gly Ala Pro Glu Asp Ser Trp Arg Arg Arg Ile Pro100 105 110 Ser Ile Phe Leu Asp Val Ala Arg Phe Ala Leu Ile Ala Val GlyIle 115 120 125 Thr Val Ile Met Ala Tyr Val Trp Gly Ala Asn Val Gly GlyLeu Phe 130 135 140 Thr Ala Leu Gly Val Thr Ser Ile Val Leu Gly Leu AlaLeu Gln Asn 145 150 155 160 Ser Val Gly Gln Ile Ile Ser Gly Leu Leu LeuLeu Phe Glu Gln Pro 165 170 175 Phe Arg Leu Gly Asp Trp Ile Thr Val ProThr Ala Ala Gly Arg Pro 180 185 190 Ser Ala His Gly Arg Val Val Glu ValAsn Trp Arg Ala Thr His Ile 195 200 205 Asp Thr Gly Gly Asn Leu Leu ValMet Pro Asn Ala Glu Leu Ala Gly 210 215 220 Ala Ser Phe Thr Asn Tyr SerArg Pro Val Gly Glu His Arg Leu Thr 225 230 235 240 Val Val Thr Thr PheAsn Ala Ala Asp Thr Pro Asp Asp Val Cys Glu 245 250 255 Met Leu Ser SerVal Ala Ala Ser Leu Pro Glu Leu Arg Thr Asp Gly 260 265 270 Gln Ile AlaThr Leu Tyr Leu Gly Ala Ala Glu Tyr Glu Lys Ser Ile 275 280 285 Pro LeuHis Thr Pro Ala Val Asp Asp Ser Val Arg Ser Thr Tyr Leu 290 295 300 ArgTrp Val Trp Tyr Ala Ala Arg Arg Gln Glu Leu Arg Xaa Asn Gly 305 310 315320 Val Ala Asp Xaa Phe Asp Thr Pro Glu Arg Ile Ala Ser Ala Met Arg 325330 335 Ala Val Ala Ser Thr Leu Arg Leu Ala Asp Asp Glu Gln Gln Glu Ile340 345 350 Ala Asp Val Val Arg Leu Val Arg Tyr Gly Asn Gly Glu Arg LeuGln 355 360 365 Gln Pro Gly Gln Val Pro Thr Gly Met Arg Phe Ile Val AspGly Arg 370 375 380 Val Ser Leu Ser Val Ile Asp Gln Asp Gly Asp Val IlePro Ala Arg 385 390 395 400 Val Leu Glu Arg Gly Asp Phe Leu Gly Gln ThrThr Leu Thr Arg Glu 405 410 415 Pro Val Leu Ala Thr Ala His Ala Leu GluGlu Val Thr Val Leu Glu 420 425 430 Met Ala Arg Asp Glu Ile Glu Arg LeuVal His Arg Lys Pro Ile Leu 435 440 445 Leu His Val Ile Gly Ala Val IleAla Asp Arg Arg Ala His Glu Leu 450 455 460 Arg Leu Met Asp Ser Gln Asp465 470 <210> SEQ ID NO 173 <211> LENGTH: 2172 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 173 tagatgacaa ttctgccctggaatgcgcga acgtctgaac acccgacgcg aaaaagacgc 60 gggcgctacc acctcctgtcgcggatgagc atccagtcca agttgctgct gatgctgctt 120 ctgaccagca ttctctcggctgcggtggtc ggtttcatcg gctatcagtc cggacggtcc 180 tcgctgcgcg catcggtgttcgaccgcctc accgacatcc gcgagtcgca gtcgcgcggg 240 ttggagaatc agttcgcggacctgaagaac tcgatggtga tttactcgcg cggcagcact 300 gccacggagg cgatcggcgcgttcagcgac ggtttccgtc agctcggcga tgcgacgatc 360 aataccgggc aggcggcgtcattgcgccgt tactacgacc ggacgttcgc caacaccacc 420 ctcgacgaca gcggaaaccgcgtcgacgtc cgcgcgctca tcccgaaatc caacccccag 480 cgctatctgc aggcgctctataccccgccg tttcagaact gggagaaggc gatcgcgttc 540 gacgacgcgc gcgacggcagcgcctggtcg gccgccaatg ccagattcaa cgagttcttc 600 cgcgagatcg tgcaccgcttcaacttcgag gatctgatgc tgctcgacct cgagggcaac 660 gtggtgtact ccgcctacaaggggccggat ctcgggacaa acatcgtcaa cggcccctat 720 cgcaaccggg aactgtcggaagcctacgag aaggcggtcg cgtcgaactc gatcgactat 780 gtcggtgtca ccgacttcgggtggtacctg cctgccgagg aaccgaccgc ctggttcctg 840 tccccggtcg ggttgaaggaccgagtcgac ggtgtgatgg cggtccagtt cccgatcgcg 900 cggatcaacg aattgatgacggcgcgggga cagtggcgtg acaccgggat gggagacacc 960 ggtgagacca tcctggtcggaccggacaat ctgatgcgct cggactcccg gctgttccgc 1020 gagaaccggg agaagttcctggccgacgtc gtcgaggggg gaaccccgcc ggaggtcgcc 1080 gacgaatcgg ttgaccgccgcggcaccacg ctggtgcagc cggtgaccac ccgctccgtc 1140 gaggaggccc aacgcggcaacaccgggacg acgatcgagg acgactatct cggccacgag 1200 gcgttacagg cgtactcaccggtggacctg ccgggactgc actgggtgat cgtggccaag 1260 atcgacaccg acgaggcgttcgccccggtg gcgcagttca ccaggaccct ggtgctgtcg 1320 acggtgatca tcatcttcggcgtgtcgctg gcggccatgc tgctggcgcg gttgttcgtc 1380 cgtccgatcc ggcggttgcaggccggcgcc cagcagatca gcggcggtga ctaccgcctc 1440 gctctgccgg tgttgtctcgtgacgaattc ggcgatctga caacagcttt caacgacatg 1500 agtcgcaatc tgtcgatcaaggacgagctg ctcggcgagg agcgcgccga gaaccaacgg 1560 ctgatgctgt ccctgatgcccgaaccggtg atgcagcgct acctcgacgg ggaggagacg 1620 atcgcccagg accacaagaacgtcacggtg atcttcgccg acatgatggg cctcgacgag 1680 ttgtcgcgca tgttgacctccgaggaactg atggtggtgg tcaacgacct gacccgccag 1740 ttcgacgccg ccgccgagagtctcggggtc gaccacgtgc ggacgctgca cgacgggtac 1800 ctggccagct gcgggttaggcgtgccgcgg ctggacaacg tccggcgcac ggtcaatttc 1860 gcgatcgaaa tggaccgcatcatcgaccgg cacgccgccg agtccgggca cgacctgcgg 1920 ctccgcgcgg gcatcgacaccgggtcggcg gccagcgggc tggtggggcg gtccacgttg 1980 gcgtacgaca tgtggggttcggcggtcgat gtcgctaacc aggtgcagcg cggctccccc 2040 cagcccggca tctacgtcacctcgcgggtg cacgaggtca tgcaggaaac tctcgacttc 2100 gtcgccgccg gggaggtcgtcggcgagcgc ggcgtcgaga cggtctggcg gttgcagggc 2160 caccggcgat ga 2172<210> SEQ ID NO 174 <211> LENGTH: 722 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 174 Met Thr Ile Leu Pro Trp Asn AlaArg Thr Ser Glu His Pro Thr Arg 1 5 10 15 Lys Arg Arg Gly Arg Tyr HisLeu Leu Ser Arg Met Ser Ile Gln Ser 20 25 30 Lys Leu Leu Leu Met Leu LeuLeu Thr Ser Ile Leu Ser Ala Ala Val 35 40 45 Val Gly Phe Ile Gly Tyr GlnSer Gly Arg Ser Ser Leu Arg Ala Ser 50 55 60 Val Phe Asp Arg Leu Thr AspIle Arg Glu Ser Gln Ser Arg Gly Leu 65 70 75 80 Glu Asn Gln Phe Ala AspLeu Lys Asn Ser Met Val Ile Tyr Ser Arg 85 90 95 Gly Ser Thr Ala Thr GluAla Ile Gly Ala Phe Ser Asp Gly Phe Arg 100 105 110 Gln Leu Gly Asp AlaThr Ile Asn Thr Gly Gln Ala Ala Ser Leu Arg 115 120 125 Arg Tyr Tyr AspArg Thr Phe Ala Asn Thr Thr Leu Asp Asp Ser Gly 130 135 140 Asn Arg ValAsp Val Arg Ala Leu Ile Pro Lys Ser Asn Pro Gln Arg 145 150 155 160 TyrLeu Gln Ala Leu Tyr Thr Pro Pro Phe Gln Asn Trp Glu Lys Ala 165 170 175Ile Ala Phe Asp Asp Ala Arg Asp Gly Ser Ala Trp Ser Ala Ala Asn 180 185190 Ala Arg Phe Asn Glu Phe Phe Arg Glu Ile Val His Arg Phe Asn Phe 195200 205 Glu Asp Leu Met Leu Leu Asp Leu Glu Gly Asn Val Val Tyr Ser Ala210 215 220 Tyr Lys Gly Pro Asp Leu Gly Thr Asn Ile Val Asn Gly Pro TyrArg 225 230 235 240 Asn Arg Glu Leu Ser Glu Ala Tyr Glu Lys Ala Val AlaSer Asn Ser 245 250 255 Ile Asp Tyr Val Gly Val Thr Asp Phe Gly Trp TyrLeu Pro Ala Glu 260 265 270 Glu Pro Thr Ala Trp Phe Leu Ser Pro Val GlyLeu Lys Asp Arg Val 275 280 285 Asp Gly Val Met Ala Val Gln Phe Pro IleAla Arg Ile Asn Glu Leu 290 295 300 Met Thr Ala Arg Gly Gln Trp Arg AspThr Gly Met Gly Asp Thr Gly 305 310 315 320 Glu Thr Ile Leu Val Gly ProAsp Asn Leu Met Arg Ser Asp Ser Arg 325 330 335 Leu Phe Arg Glu Asn ArgGlu Lys Phe Leu Ala Asp Val Val Glu Gly 340 345 350 Gly Thr Pro Pro GluVal Ala Asp Glu Ser Val Asp Arg Arg Gly Thr 355 360 365 Thr Leu Val GlnPro Val Thr Thr Arg Ser Val Glu Glu Ala Gln Arg 370 375 380 Gly Asn ThrGly Thr Thr Ile Glu Asp Asp Tyr Leu Gly His Glu Ala 385 390 395 400 LeuGln Ala Tyr Ser Pro Val Asp Leu Pro Gly Leu His Trp Val Ile 405 410 415Val Ala Lys Ile Asp Thr Asp Glu Ala Phe Ala Pro Val Ala Gln Phe 420 425430 Thr Arg Thr Leu Val Leu Ser Thr Val Ile Ile Ile Phe Gly Val Ser 435440 445 Leu Ala Ala Met Leu Leu Ala Arg Leu Phe Val Arg Pro Ile Arg Arg450 455 460 Leu Gln Ala Gly Ala Gln Gln Ile Ser Gly Gly Asp Tyr Arg LeuAla 465 470 475 480 Leu Pro Val Leu Ser Arg Asp Glu Phe Gly Asp Leu ThrThr Ala Phe 485 490 495 Asn Asp Met Ser Arg Asn Leu Ser Ile Lys Asp GluLeu Leu Gly Glu 500 505 510 Glu Arg Ala Glu Asn Gln Arg Leu Met Leu SerLeu Met Pro Glu Pro 515 520 525 Val Met Gln Arg Tyr Leu Asp Gly Glu GluThr Ile Ala Gln Asp His 530 535 540 Lys Asn Val Thr Val Ile Phe Ala AspMet Met Gly Leu Asp Glu Leu 545 550 555 560 Ser Arg Met Leu Thr Ser GluGlu Leu Met Val Val Val Asn Asp Leu 565 570 575 Thr Arg Gln Phe Asp AlaAla Ala Glu Ser Leu Gly Val Asp His Val 580 585 590 Arg Thr Leu His AspGly Tyr Leu Ala Ser Cys Gly Leu Gly Val Pro 595 600 605 Arg Leu Asp AsnVal Arg Arg Thr Val Asn Phe Ala Ile Glu Met Asp 610 615 620 Arg Ile IleAsp Arg His Ala Ala Glu Ser Gly His Asp Leu Arg Leu 625 630 635 640 ArgAla Gly Ile Asp Thr Gly Ser Ala Ala Ser Gly Leu Val Gly Arg 645 650 655Ser Thr Leu Ala Tyr Asp Met Trp Gly Ser Ala Val Asp Val Ala Asn 660 665670 Gln Val Gln Arg Gly Ser Pro Gln Pro Gly Ile Tyr Val Thr Ser Arg 675680 685 Val His Glu Val Met Gln Glu Thr Leu Asp Phe Val Ala Ala Gly Glu690 695 700 Val Val Gly Glu Arg Gly Val Glu Thr Val Trp Arg Leu Gln GlyHis 705 710 715 720 Arg Arg <210> SEQ ID NO 175 <211> LENGTH: 898 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 175gagcaaccgt tccggctcgg cgactggatc accgtcccca ccgcggcggg ccggccgtcc 60gcccacggcc gcgtggtgga agtcaactgg cgtgcaacac atatcgacac cggcggcaac 120ctgctggtaa tgcccaacgc cgaactcgcc ggcgcgtcgt tcaccaatta cagccggccc 180gtgggagagc accggctgac cgtcgtcacc accttcaacg ccgcggacac ccccgatgat 240gtctgcgaga tgctgtcgtc ggtcgcggcg tcgctgcccg aactgcgcac cgacggacag 300atcgccacgc tctatctcgg tgcggccgaa tacgagaagt cgatcccgtt gcacacaccc 360gcggtggacg actcggtcag gagcacgtac ctgcgatggg tctggtacgc cgcgcgccgg 420caggaacttc gcctaacggc gtcgccgacg attcgacacg ccggaacgga tcgcctcggc 480catgcgggct gtggcgtcca cactgcgctt ggcagacgac gaacagcagg agatcgccga 540cgtggtgcgt ctggtccgtt acggcaacgg ggaacgcctc cagcagccgg gtcaggtacc 600gaccgggatg aggttcatcg tagacggcag ggtgagtctg tccgtgatcg atcaggacgg 660cgacgtgatc ccggcgcggg tgctcgagcg tggcgacttc ctggggcaga ccacgctgac 720gcgggaaccg gtactggcga ccgcgcacgc gctggaggaa gtcaccgtgc tggagatggc 780ccgtgacgag atcgagcgcc tggtgcaccg aaagccgatc ctgctgcacg tgatcggggc 840cgtgatcgcc gaccggcgcg cgcacgaact tcggttgatg gcggactcgc aggactga 898<210> SEQ ID NO 176 <211> LENGTH: 2013 <212> TYPE: DNA <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 176 ggctatcagt ccggacggtcctcgctgcgc gcatcggtgt tcgaccgcct caccgacatc 60 cgcgagtcgc agtcgcgcgggttggagaat cagttcgcgg acctgaagaa ctcgatggtg 120 atttactcgc gcggcagcactgccacggag gcgatcggcg cgttcagcga cggtttccgt 180 cagctcggcg atgcgacgatcaataccggg caggcggcgt cattgcgccg ttactacgac 240 cggacgttcg ccaacaccaccctcgacgac agcggaaacc gcgtcgacgt ccgcgcgctc 300 atcccgaaat ccaacccccagcgctatctg caggcgctct ataccccgcc gtttcagaac 360 tgggagaagg cgatcgcgttcgacgacgcg cgcgacggca gcgcctggtc ggccgccaat 420 gccagattca acgagttcttccgcgagatc gtgcaccgct tcaacttcga ggatctgatg 480 ctgctcgacc tcgagggcaacgtggtgtac tccgcctaca aggggccgga tctcgggaca 540 aacatcgtca acggcccctatcgcaaccgg gaactgtcgg aagcctacga gaaggcggtc 600 gcgtcgaact cgatcgactatgtcggtgtc accgacttcg ggtggtacct gcctgccgag 660 gaaccgaccg cctggttcctgtccccggtc gggttgaagg accgagtcga cggtgtgatg 720 gcggtccagt tcccgatcgcgcggatcaac gaattgatga cggcgcgggg acagtggcgt 780 gacaccggga tgggagacaccggtgagacc atcctggtcg gaccggacaa tctgatgcgc 840 tcggactccc ggctgttccgcgagaaccgg gagaagttcc tggccgacgt cgtcgagggg 900 ggaaccccgc cggaggtcgccgacgaatcg gttgaccgcc gcggcaccac gctggtgcag 960 ccggtgacca cccgctccgtcgaggaggcc caacgcggca acaccgggac gacgatcgag 1020 gacgactatc tcggccacgaggcgttacag gcgtactcac cggtggacct gccgggactg 1080 cactgggtga tcgtggccaagatcgacacc gacgaggcgt tcgccccggt ggcgcagttc 1140 accaggaccc tggtgctgtcgacggtgatc atcatcttcg gcgtgtcgct ggcggccatg 1200 ctgctggcgc ggttgttcgtccgtccgatc cggcggttgc aggccggcgc ccagcagatc 1260 agcggcggtg actaccgcctcgctctgccg gtgttgtctc gtgacgaatt cggcgatctg 1320 acaacagctt tcaacgacatgagtcgcaat ctgtcgatca aggacgagct gctcggcgag 1380 gagcgcgccg agaaccaacggctgatgctg tccctgatgc ccgaaccggt gatgcagcgc 1440 tacctcgacg gggaggagacgatcgcccag gaccacaaga acgtcacggt gatcttcgcc 1500 gacatgatgg gcctcgacgagttgtcgcgc atgttgacct ccgaggaact gatggtggtg 1560 gtcaacgacc tgacccgccagttcgacgcc gccgccgaga gtctcggggt cgaccacgtg 1620 cggacgctgc acgacgggtacctggccagc tgcgggttag gcgtgccgcg gctggacaac 1680 gtccggcgca cggtcaatttcgcgatcgaa atggaccgca tcatcgaccg gcacgccgcc 1740 gagtccgggc acgacctgcggctccgcgcg ggcatcgaca ccgggtcggc ggccagcggg 1800 ctggtggggc ggtccacgttggcgtacgac atgtggggtt cggcggtcga tgtcgctaac 1860 caggtgcagc gcggctccccccagcccggc atctacgtca cctcgcgggt gcacgaggtc 1920 atgcaggaaa ctctcgacttcgtcgccgcc ggggaggtcg tcggcgagcg cggcgtcgag 1980 acggtctggc ggttgcagggccaccggcga tga 2013 <210> SEQ ID NO 177 <211> LENGTH: 297 <212> TYPE:PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:UNSURE <222> LOCATION: (145)...(145) <221> NAME/KEY: UNSURE <222>LOCATION: (151)...(151) <400> SEQUENCE: 177 Glu Gln Pro Phe Arg Leu GlyAsp Trp Ile Thr Val Pro Thr Ala Ala 1 5 10 15 Gly Arg Pro Ser Ala HisGly Arg Val Val Glu Val Asn Trp Arg Ala 20 25 30 Thr His Ile Asp Thr GlyGly Asn Leu Leu Val Met Pro Asn Ala Glu 35 40 45 Leu Ala Gly Ala Ser PheThr Asn Tyr Ser Arg Pro Val Gly Glu His 50 55 60 Arg Leu Thr Val Val ThrThr Phe Asn Ala Ala Asp Thr Pro Asp Asp 65 70 75 80 Val Cys Glu Met LeuSer Ser Val Ala Ala Ser Leu Pro Glu Leu Arg 85 90 95 Thr Asp Gly Gln IleAla Thr Leu Tyr Leu Gly Ala Ala Glu Tyr Glu 100 105 110 Lys Ser Ile ProLeu His Thr Pro Ala Val Asp Asp Ser Val Arg Ser 115 120 125 Thr Tyr LeuArg Trp Val Trp Tyr Ala Ala Arg Arg Gln Glu Leu Arg 130 135 140 Xaa AsnGly Val Ala Asp Xaa Phe Asp Thr Pro Glu Arg Ile Ala Ser 145 150 155 160Ala Met Arg Ala Val Ala Ser Thr Leu Arg Leu Ala Asp Asp Glu Gln 165 170175 Gln Glu Ile Ala Asp Val Val Arg Leu Val Arg Tyr Gly Asn Gly Glu 180185 190 Arg Leu Gln Gln Pro Gly Gln Val Pro Thr Gly Met Arg Phe Ile Val195 200 205 Asp Gly Arg Val Ser Leu Ser Val Ile Asp Gln Asp Gly Asp ValIle 210 215 220 Pro Ala Arg Val Leu Glu Arg Gly Asp Phe Leu Gly Gln ThrThr Leu 225 230 235 240 Thr Arg Glu Pro Val Leu Ala Thr Ala His Ala LeuGlu Glu Val Thr 245 250 255 Val Leu Glu Met Ala Arg Asp Glu Ile Glu ArgLeu Val His Arg Lys 260 265 270 Pro Ile Leu Leu His Val Ile Gly Ala ValAla Asp Arg Arg Ala His 275 280 285 Glu Leu Arg Leu Met Asp Ser Gln Asp290 295 <210> SEQ ID NO 178 <211> LENGTH: 670 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 178 Gly Tyr Gln Ser GlyArg Ser Ser Leu Arg Ala Ser Val Phe Asp Arg 1 5 10 15 Leu Thr Asp IleArg Glu Ser Gln Ser Arg Gly Leu Glu Asn Gln Phe 20 25 30 Ala Asp Leu LysAsn Ser Met Val Ile Tyr Ser Arg Gly Ser Thr Ala 35 40 45 Thr Glu Ala IleGly Ala Phe Ser Asp Gly Phe Arg Gln Leu Gly Asp 50 55 60 Ala Thr Ile AsnThr Gly Gln Ala Ala Ser Leu Arg Arg Tyr Tyr Asp 65 70 75 80 Arg Thr PheAla Asn Thr Thr Leu Asp Asp Ser Gly Asn Arg Val Asp 85 90 95 Val Arg AlaLeu Ile Pro Lys Ser Asn Pro Gln Arg Tyr Leu Gln Ala 100 105 110 Leu TyrThr Pro Pro Phe Gln Asn Trp Glu Lys Ala Ile Ala Phe Asp 115 120 125 AspAla Arg Asp Gly Ser Ala Trp Ser Ala Ala Asn Ala Arg Phe Asn 130 135 140Glu Phe Phe Arg Glu Ile Val His Arg Phe Asn Phe Glu Asp Leu Met 145 150155 160 Leu Leu Asp Leu Glu Gly Asn Val Val Tyr Ser Ala Tyr Lys Gly Pro165 170 175 Asp Leu Gly Thr Asn Ile Val Asn Gly Pro Tyr Arg Asn Arg GluLeu 180 185 190 Ser Glu Ala Tyr Glu Lys Ala Val Ala Ser Asn Ser Ile AspTyr Val 195 200 205 Gly Val Thr Asp Phe Gly Trp Tyr Leu Pro Ala Glu GluPro Thr Ala 210 215 220 Trp Phe Leu Ser Pro Val Gly Leu Lys Asp Arg ValAsp Gly Val Met 225 230 235 240 Ala Val Gln Phe Pro Ile Ala Arg Ile AsnGlu Leu Met Thr Ala Arg 245 250 255 Gly Gln Trp Arg Asp Thr Gly Met GlyAsp Thr Gly Glu Thr Ile Leu 260 265 270 Val Gly Pro Asp Asn Leu Met ArgSer Asp Ser Arg Leu Phe Arg Glu 275 280 285 Asn Arg Glu Lys Phe Leu AlaAsp Val Val Glu Gly Gly Thr Pro Pro 290 295 300 Glu Val Ala Asp Glu SerVal Asp Arg Arg Gly Thr Thr Leu Val Gln 305 310 315 320 Pro Val Thr ThrArg Ser Val Glu Glu Ala Gln Arg Gly Asn Thr Gly 325 330 335 Thr Thr IleGlu Asp Asp Tyr Leu Gly His Glu Ala Leu Gln Ala Tyr 340 345 350 Ser ProVal Asp Leu Pro Gly Leu His Trp Val Ile Val Ala Lys Ile 355 360 365 AspThr Asp Glu Ala Phe Ala Pro Val Ala Gln Phe Thr Arg Thr Leu 370 375 380Val Leu Ser Thr Val Ile Ile Ile Phe Gly Val Ser Leu Ala Ala Met 385 390395 400 Leu Leu Ala Arg Leu Phe Val Arg Pro Ile Arg Arg Leu Gln Ala Gly405 410 415 Ala Gln Gln Ile Ser Gly Gly Asp Tyr Arg Leu Ala Leu Pro ValLeu 420 425 430 Ser Arg Asp Glu Phe Gly Asp Leu Thr Thr Ala Phe Asn AspMet Ser 435 440 445 Arg Asn Leu Ser Ile Lys Asp Glu Leu Leu Gly Glu GluArg Ala Glu 450 455 460 Asn Gln Arg Leu Met Leu Ser Leu Met Pro Glu ProVal Met Gln Arg 465 470 475 480 Tyr Leu Asp Gly Glu Glu Thr Ile Ala GlnAsp His Lys Asn Val Thr 485 490 495 Val Ile Phe Ala Asp Met Met Gly LeuAsp Glu Leu Ser Arg Met Leu 500 505 510 Thr Ser Glu Glu Leu Met Val ValVal Asn Asp Leu Thr Arg Gln Phe 515 520 525 Asp Ala Ala Ala Glu Ser LeuGly Val Asp His Val Arg Thr Leu His 530 535 540 Asp Gly Tyr Leu Ala SerCys Gly Leu Gly Val Pro Arg Leu Asp Asn 545 550 555 560 Val Arg Arg ThrVal Asn Phe Ala Ile Glu Met Asp Arg Ile Ile Asp 565 570 575 Arg His AlaAla Glu Ser Gly His Asp Leu Arg Leu Arg Ala Gly Ile 580 585 590 Asp ThrGly Ser Ala Ala Ser Gly Leu Val Gly Arg Ser Thr Leu Ala 595 600 605 TyrAsp Met Trp Gly Ser Ala Val Asp Val Ala Asn Gln Val Gln Arg 610 615 620Gly Ser Pro Gln Pro Gly Ile Tyr Val Thr Ser Arg Val His Glu Val 625 630635 640 Met Gln Glu Thr Leu Asp Phe Val Ala Ala Gly Glu Val Val Gly Glu645 650 655 Arg Gly Val Glu Thr Val Trp Arg Leu Gln Gly His Arg Arg 660665 670 <210> SEQ ID NO 179 <211> LENGTH: 520 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 179 gtgatcgacg aaaccctcttccatgccgag gagaagatgg agaaggccgt ctcggtggca 60 cccgacgacc tggcgtcgattcgtaccggc cgcgcgaacc ccggcatgtt caaccggatc 120 aacatcgact actacggcgcctccaccccg atcacgcagc tgtccagcat caacgtgccc 180 gaggcgcgca tggtggtgatcaagccctac gaggcgagcc agctgcgcct catcgaggat 240 gcgatccgca actccgacctcggcgtcaat ccgaccaacg acggcaacat catccgggtg 300 tcgatcccgc agctcaccgaggagcgccgc cgcgacctgg tcaagcaggc caaggccaag 360 ggcgaggacg ccaaggtgtcggtgcgcaac atccgtcgca acgatatgaa cacctttcgc 420 atcgcaccgg tacggctgccgacgccaccg ccgtcgtaga agcgacagag gatcgcaggt 480 aacggtattg gccacgccttctgtggcggg ccgacaccac 520 <210> SEQ ID NO 180 <211> LENGTH: 1071 <212>TYPE: DNA <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 180cgtggggaag gattgcactc tatgagcgaa atcgcccgtc cctggcgggt tctggcaggt 60ggcatcggtg cctgcgccgc gggtatcgcc ggggtgctga gcatcgcggt caccacggcg 120tcggcccagc cgggcctccc gcagcccccg ctgcccgccc ctgccacagt gacgcaaacc 180gtcacggttg cgcccaacgc cgcgccacaa ctcatcccgc gccccggtgt gacgcctgcc 240accggcggcg ccgccgcggt gcccgccggg gtgagcgccc cggcggtcgc gccggccccc 300gcgctgcccg cccgcccggt gtccacgatc gccccggcca cctcgggcac gctcagcgag 360ttcttcgccg ccaagggcgt cacgatggag ccgcagtcca gccgcgactt ccgcgccctc 420aacatcgtgc tgccgaagcc gcggggctgg gagcacatcc cggacccgaa cgtgccggac 480gcgttcgcgg tgctggccga ccgggtcggc ggcaacggcc tgtactcgtc gaacgcccag 540gtggtggtct acaaactcgt cggcgagttc gaccccaagg aagcgatcag ccacggcttc 600gtcgacagcc agaagctgcc ggcgtggcgt tccaccgacg cgtcgctggc cgacttcggc 660ggaatgccgt cctcgctgat cgagggcacc taccgcgaga acaacatgaa gctgaacacg 720tcccggcgcc acgtcattgc caccgcgggg cccgaccact acctggtgtc gctgtcggtg 780accaccagcg tcgaacaggc cgtggccgaa gccgcggagg ccaccgacgc gattgtcaac 840ggcttcaagg tcagcgttcc gggtccgggt ccggccgcac cgccacctgc acccggtgcc 900cccggtgtcc cgcccgcccc cggcgccccg gcgctgccgc tggccgtcgc accacccccg 960gctcccgctg ttcccgccgt ggcgcccgcg ccacagctgc tgggactgca gggatagacg 1020tcgtcgtccc ccgggcgaag cctggcgccc gggggacgac ggcccctttc t 1071 <210> SEQID NO 181 <211> LENGTH: 152 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <400> SEQUENCE: 181 Val Ile Asp Glu Thr Leu Phe HisAla Glu Glu Lys Met Glu Lys Ala 1 5 10 15 Val Ser Val Ala Pro Asp AspLeu Ala Ser Ile Arg Thr Gly Arg Ala 20 25 30 Asn Pro Gly Met Phe Asn ArgIle Asn Ile Asp Tyr Tyr Gly Ala Ser 35 40 45 Thr Pro Ile Thr Gln Leu SerSer Ile Asn Val Pro Glu Ala Arg Met 50 55 60 Val Val Ile Lys Pro Tyr GluAla Ser Gln Leu Arg Leu Ile Glu Asp 65 70 75 80 Ala Ile Arg Asn Ser AspLeu Gly Val Asn Pro Thr Asn Asp Gly Asn 85 90 95 Ile Ile Arg Val Ser IlePro Gln Leu Thr Glu Glu Arg Arg Arg Asp 100 105 110 Leu Val Lys Gln AlaLys Ala Lys Gly Glu Asp Ala Lys Val Ser Val 115 120 125 Arg Asn Ile ArgArg Asn Asp Met Asn Thr Phe Arg Ile Ala Pro Val 130 135 140 Arg Leu ProThr Pro Pro Pro Ser 145 150 <210> SEQ ID NO 182 <211> LENGTH: 331 <212>TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 182 MetSer Glu Ile Ala Arg Pro Trp Arg Val Leu Ala Gly Gly Ile Gly 1 5 10 15Ala Cys Ala Ala Gly Ile Ala Gly Val Leu Ser Ile Ala Val Thr Thr 20 25 30Ala Ser Ala Gln Pro Gly Leu Pro Gln Pro Pro Leu Pro Ala Pro Ala 35 40 45Thr Val Thr Gln Thr Val Thr Val Ala Pro Asn Ala Ala Pro Gln Leu 50 55 60Ile Pro Arg Pro Gly Val Thr Pro Ala Thr Gly Gly Ala Ala Ala Val 65 70 7580 Pro Ala Gly Val Ser Ala Pro Ala Val Ala Pro Ala Pro Ala Leu Pro 85 9095 Ala Arg Pro Val Ser Thr Ile Ala Pro Ala Thr Ser Gly Thr Leu Ser 100105 110 Glu Phe Phe Ala Ala Lys Gly Val Thr Met Glu Pro Gln Ser Ser Arg115 120 125 Asp Phe Arg Ala Leu Asn Ile Val Leu Pro Lys Pro Arg Gly TrpGlu 130 135 140 His Ile Pro Asp Pro Asn Val Pro Asp Ala Phe Ala Val LeuAla Asp 145 150 155 160 Arg Val Gly Gly Asn Gly Leu Tyr Ser Ser Asn AlaGln Val Val Val 165 170 175 Tyr Lys Leu Val Gly Glu Phe Asp Pro Lys GluAla Ile Ser His Gly 180 185 190 Phe Val Asp Ser Gln Lys Leu Pro Ala TrpArg Ser Thr Asp Ala Ser 195 200 205 Leu Ala Asp Phe Gly Gly Met Pro SerSer Leu Ile Glu Gly Thr Tyr 210 215 220 Arg Glu Asn Asn Met Lys Leu AsnThr Ser Arg Arg His Val Ile Ala 225 230 235 240 Thr Ala Gly Pro Asp HisTyr Leu Val Ser Leu Ser Val Thr Thr Ser 245 250 255 Val Glu Gln Ala ValAla Glu Ala Ala Glu Ala Thr Asp Ala Ile Val 260 265 270 Asn Gly Phe LysVal Ser Val Pro Gly Pro Gly Pro Ala Ala Pro Pro 275 280 285 Pro Ala ProGly Ala Pro Gly Val Pro Pro Ala Pro Gly Ala Pro Ala 290 295 300 Leu ProLeu Ala Val Ala Pro Pro Pro Ala Pro Ala Val Pro Ala Val 305 310 315 320Ala Pro Ala Pro Gln Leu Leu Gly Leu Gln Gly 325 330 <210> SEQ ID NO 183<211> LENGTH: 207 <212> TYPE: DNA <213> ORGANISM: Mycobacterium vaccae<400> SEQUENCE: 183 acctacgagt tcgagaacaa ggtcacgggc ggccgcatcccgcgcgagta catcccgtcg 60 gtggatgccg gcgcgcagga cgccatgcag tacggcgtgctggccggcta cccgctggtt 120 aacgtcaagc tgacgctgct cgacggtgcc taccacgaagtcgactcgtc ggaaatggca 180 ttcaaggttg ccggctccca ggtcata 207 <210> SEQ IDNO 184 <211> LENGTH: 69 <212> TYPE: PRT <213> ORGANISM: Mycobacteriumvaccae <400> SEQUENCE: 184 Thr Tyr Glu Phe Glu Asn Lys Val Thr Gly GlyArg Ile Pro Arg Glu 1 5 10 15 Tyr Ile Pro Ser Val Asp Ala Gly Ala GlnAsp Ala Met Gln Tyr Gly 20 25 30 Val Leu Ala Gly Tyr Pro Leu Val Asn ValLys Leu Thr Leu Leu Asp 35 40 45 Gly Ala Tyr His Glu Val Asp Ser Ser GluMet Ala Phe Lys Val Ala 50 55 60 Gly Ser Gln Val Ile 65 <210> SEQ ID NO185 <211> LENGTH: 898 <212> TYPE: DNA <213> ORGANISM: Mycobacteriumvaccae <220> FEATURE: <221> NAME/KEY: unsure <222> LOCATION:(637)...(637) <221> NAME/KEY: unsure <222> LOCATION: (662)...(662) <400>SEQUENCE: 185 cgacctccac ccgggcgtga ggccaaccac taggctggtc accagtagtcgacggcacac 60 ttcaccgaaa aaatgaggac agaggagaca cccgtgacga tccgtgttggtgtgaacggc 120 ttcggccgta tcggacgcaa cttcttccgc gcgctggacg cgcagaaggccgaaggcaag 180 aacaaggaca tcgagatcgt cgcggtcaac gacctcaccg acaacgccacgctggcgcac 240 ctgctgaagt tcgactcgat cctgggccgg ctgccctacg acgtgagcctcgaaggcgag 300 gacaccatcg tcgtcggcag caccaagatc aaggcgctcg aggtcaaggaaggcccggcg 360 gcgctgccct ggggcgacct gggcgtcgac gtcgtcgtcg agtccaccggcatcttcacc 420 aagcgcgaca aggcccaggg ccacctcgac gcgggcgcca agaaggtcatcatctccgcg 480 ccggccaccg atgaggacat caccatcgtg ctcggcgtca acgacgacaagtacgacggc 540 agccagaaca tcatctccaa cgcgtcgtgc accacgaact gcctcggcccgctggcgaag 600 gtcatcaacg acgagttcgg catcgtcaag ggcctgntga ccaccatccacgcctacacc 660 cnggtccaga acctgcagga cggcccgcac aaggatctgc gccgggcccgcgccgccgcg 720 ctgaacatcg tgccgacctc caccggtgcc gccaaggcca tcggactggtgctgcccgag 780 ctgaagggca agctcgacgg ctacgcgctg cgggtgccga tccccaccggctcggtcacc 840 gacctgaccg ccgagctggg caagtcggcc accgtggacg agatcaacgccgcgatga 898 <210> SEQ ID NO 186 <211> LENGTH: 268 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE<222> LOCATION: (182)...(182) <221> NAME/KEY: UNSURE <222> LOCATION:(190)...(190) <400> SEQUENCE: 186 Val Thr Ile Arg Val Gly Val Asn GlyPhe Gly Arg Ile Gly Arg Asn 1 5 10 15 Phe Phe Arg Ala Leu Asp Ala GlnLys Ala Glu Gly Lys Asn Lys Asp 20 25 30 Ile Glu Ile Val Ala Val Asn AspLeu Thr Asp Asn Ala Thr Leu Ala 35 40 45 His Leu Leu Lys Phe Asp Ser IleLeu Gly Arg Leu Pro Tyr Asp Val 50 55 60 Ser Leu Glu Gly Glu Asp Thr IleVal Val Gly Ser Thr Lys Ile Lys 65 70 75 80 Ala Leu Glu Val Lys Glu GlyPro Ala Ala Leu Pro Trp Gly Asp Leu 85 90 95 Gly Val Asp Val Val Val GluSer Thr Gly Ile Phe Thr Lys Arg Asp 100 105 110 Lys Ala Gln Gly His LeuAsp Ala Gly Ala Lys Lys Val Ile Ile Ser 115 120 125 Ala Pro Ala Thr AspGlu Asp Ile Thr Ile Val Leu Gly Val Asn Asp 130 135 140 Asp Lys Tyr AspGly Ser Gln Asn Ile Ile Ser Asn Ala Ser Cys Thr 145 150 155 160 Thr AsnCys Leu Gly Pro Leu Ala Lys Val Ile Asn Asp Glu Phe Gly 165 170 175 IleVal Lys Gly Leu Xaa Thr Thr Ile His Ala Tyr Thr Xaa Val Gln 180 185 190Asn Leu Gln Asp Gly Pro His Lys Asp Leu Arg Arg Ala Arg Ala Ala 195 200205 Ala Leu Asn Ile Val Pro Thr Ser Thr Gly Ala Ala Lys Ala Ile Gly 210215 220 Leu Val Leu Pro Glu Leu Lys Gly Lys Leu Asp Gly Tyr Ala Leu Arg225 230 235 240 Val Pro Ile Pro Thr Gly Ser Val Thr Asp Leu Thr Ala GluLeu Gly 245 250 255 Lys Ser Ala Thr Val Asp Glu Ile Asn Ala Ala Met 260265 <210> SEQ ID NO 187 <211> LENGTH: 41 <212> TYPE: PRT <213> ORGANISM:Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY: UNSURE <222>LOCATION: (39)...(39) <400> SEQUENCE: 187 Met Asn Lys Ala Glu Leu IleAsp Val Leu Thr Glu Lys Leu Gly Ser 1 5 10 15 Asp Arg Arg Gln Ala ThrAla Ala Val Glu Asn Val Val Asp Thr Ile 20 25 30 Val Ala Ala Val Pro LysXaa Val Val 35 40 <210> SEQ ID NO 188 <211> LENGTH: 26 <212> TYPE: DNA<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Made in a lab <221> NAME/KEY: unsure <222> LOCATION:(12)...(12) <400> SEQUENCE: 188 atgaayaarg cngarctsat ygaygt 26 <210>SEQ ID NO 189 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM:Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Made in alab <400> SEQUENCE: 189 atsgtrtgva cvacgttytc 20 <210> SEQ ID NO 190<211> LENGTH: 84 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence<220> FEATURE: <223> OTHER INFORMATION: Made in a lab <221> NAME/KEY:unsure <222> LOCATION: (2)...(2) <400> SEQUENCE: 190 gnactcattgacgtactcac tgagaagctg ggctcggatt gtcggcaagc gactgcggca 60 atggagaacgtggtccacac cata 84 <210> SEQ ID NO 191 <211> LENGTH: 337 <212> TYPE: DNA<213> ORGANISM: Mycobacterium vaccae <220> FEATURE: <221> NAME/KEY:unsure <222> LOCATION: (2)...(2) <400> SEQUENCE: 191 gnactcattgacgtactcac tgagaagctg ggctcggatt gtcggcaagc gactgcggcg 60 gtggagaatgttgtcgacac catcgtgcgc gccgtgcaca agggtgagag cgtcaccatc 120 acgggcttcggtgttttcga gcagcgtcgt cgcgcagcac gcgtggcacg caatccgcgc 180 accggcgagaccgtgaaggt caagcccacc tcagtcccgg cattccgtcc cggcgctcag 240 ttcaaggctgttgtctctgg cgcacagaag cttccggccg agggtccggc ggtcaagcgc 300 ggtgtgaccgcgacgagcac cgcccgcaag gcagcca 337 <210> SEQ ID NO 192 <211> LENGTH: 111<212> TYPE: PRT <213> ORGANISM: Mycobacterium vaccae <220> FEATURE:<221> NAME/KEY: UNSURE <222> LOCATION: (1)...(1) <400> SEQUENCE: 192 XaaLeu Ile Asp Val Leu Thr Glu Lys Leu Gly Ser Asp Arg Gln Ala 1 5 10 15Thr Ala Ala Val Glu Asn Val Val Asp Thr Ile Val Arg Ala Val His 20 25 30Lys Gly Glu Ser Val Thr Ile Thr Gly Phe Gly Val Phe Glu Gln Arg 35 40 45Arg Arg Ala Ala Arg Val Ala Arg Asn Pro Arg Thr Gly Glu Thr Val 50 55 60Lys Val Lys Pro Thr Ser Val Pro Ala Phe Arg Pro Gly Ala Gln Phe 65 70 7580 Lys Ala Val Val Ser Gly Ala Gln Lys Leu Pro Ala Glu Gly Pro Ala 85 9095 Val Lys Arg Gly Val Thr Ala Thr Ser Thr Ala Arg Lys Ala Ala 100 105110 <210> SEQ ID NO 193 <211> LENGTH: 1164 <212> TYPE: DNA <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 193 ggtggcgcgc atcgagaagcgcccgccccg gttcacgggc gcctgatcat ggtgcgggcg 60 gcgctgcgct acggcttcgggacggcctca ctgctggccg gcgggttcgt gctgcgcgcc 120 ctgcagggca cgcctgccgccctcggcgcg actccgggcg aggtcgcgcc ggtggcgcgc 180 cgctcgccga actaccgcgacggcaagttc gtcaacctgg agcccccgtc gggcatcacg 240 atggatcgcg acctgcagcggatgctgttg cgcgatctgg ccaacgccgc atcccagggc 300 aagccgcccg gaccgatcccgctggccgag ccgccgaagg gggatcccac tcccgcgccg 360 gcggcggcca gctggtacggccattccagc gtgctgatcg aggtcgacgg ctaccgcgtg 420 ctggccgacc cggtgtggagcaacagatgt tcgccctcac gggcggtcgg accgcagcgc 480 atgcacgacg tcccggtgccgctggaggcg cttcccgccg tggacgcggt ggtgatcagc 540 cacgaccact acgaccacctcgacatcgac accatcgtcg cgttggcgca cacccagcgg 600 gccccgttcg tggtgccgttgggcatcggc gcacacctgc gcaagtgggg cgtccccgag 660 gcgcggatcg tcgagttggactggcacgaa gcccaccgca tagacgacct gacgctggtc 720 tgcacccccg cccggcacttctccggacgg ttgttctccc gcgactcgac gctgtgggcg 780 tcgtgggtgg tcaccggctcgtcgcacaag gcgttcttcg gtggcgacac cggatacacg 840 aagagcttcg ccgagatcggcgacgagtac ggtccgttcg atctgaccct gctgccgatc 900 ggggcctacc atcccgcgttcgccgacatc cacatgaacc ccgaggaggc ggtgcgcgcc 960 catctggacc tgaccgaggtggacaacagc ctgatggtgc ccatccactg ggcgacattc 1020 cgcctcgccc cgcatccgtggtccgagccc gccgaacgcc tgctgaccgc tgccgacgcc 1080 gagcgggtac gcctgaccgtgccgattccc ggtcagcggg tggacccgga gtcgacgttc 1140 gacccgtggt ggcggttctgaacc 1164 <210> SEQ ID NO 194 <211> LENGTH: 370 <212> TYPE: PRT <213>ORGANISM: Mycobacterium vaccae <400> SEQUENCE: 194 Met Val Arg Ala AlaLeu Arg Tyr Gly Phe Gly Thr Ala Ser Leu Leu 1 5 10 15 Ala Gly Gly PheVal Leu Arg Ala Leu Gln Gly Thr Pro Ala Ala Leu 20 25 30 Gly Ala Thr ProGly Glu Val Ala Pro Val Ala Arg Arg Ser Pro Asn 35 40 45 Tyr Arg Asp GlyLys Phe Val Asn Leu Glu Pro Pro Ser Gly Ile Thr 50 55 60 Met Asp Arg AspLeu Gln Arg Met Leu Leu Arg Asp Leu Ala Asn Ala 65 70 75 80 Ala Ser GlnGly Lys Pro Pro Gly Pro Ile Pro Leu Ala Glu Pro Pro 85 90 95 Lys Gly AspPro Thr Pro Ala Pro Ala Ala Ala Ser Trp Tyr Gly His 100 105 110 Ser SerVal Leu Ile Glu Val Asp Gly Tyr Arg Val Leu Ala Asp Pro 115 120 125 ValTrp Ser Asn Arg Cys Ser Pro Ser Arg Ala Val Gly Pro Gln Arg 130 135 140Met His Asp Val Pro Val Pro Leu Glu Ala Leu Pro Ala Val Asp Ala 145 150155 160 Val Val Ile Ser His Asp His Tyr Asp His Leu Asp Ile Asp Thr Ile165 170 175 Val Ala Leu Ala His Thr Gln Arg Ala Pro Phe Val Val Pro LeuGly 180 185 190 Ile Gly Ala His Leu Arg Lys Trp Gly Val Pro Glu Ala ArgIle Val 195 200 205 Glu Leu Asp Trp His Glu Ala His Arg Ile Asp Asp LeuThr Leu Val 210 215 220 Cys Thr Pro Ala Arg His Phe Ser Gly Arg Leu PheSer Arg Asp Ser 225 230 235 240 Thr Leu Trp Ala Ser Trp Val Val Thr GlySer Ser His Lys Ala Phe 245 250 255 Phe Gly Gly Asp Thr Gly Tyr Thr LysSer Phe Ala Glu Ile Gly Asp 260 265 270 Glu Tyr Gly Pro Phe Asp Leu ThrLeu Leu Pro Ile Gly Ala Tyr His 275 280 285 Pro Ala Phe Ala Asp Ile HisMet Asn Pro Glu Glu Ala Val Arg Ala 290 295 300 His Leu Asp Leu Thr GluVal Asp Asn Ser Leu Met Val Pro Ile His 305 310 315 320 Trp Ala Thr PheArg Leu Ala Pro His Pro Trp Ser Glu Pro Ala Glu 325 330 335 Arg Leu LeuThr Ala Ala Asp Ala Glu Arg Val Arg Leu Thr Val Pro 340 345 350 Ile ProGly Gln Arg Val Asp Pro Glu Ser Thr Phe Asp Pro Trp Trp 355 360 365 ArgPhe 370

We claim:
 1. A method for the treatment of a skin disorder, comprisingadministering a composition comprising at least one component selectedfrom the group consisting of: (a) inactivated M. vaccae cells; (b) M.vaccae culture filtrate; and (c) delipidated and deglycolipidated M.vaccae cells.
 2. The method of claim 1, wherein the skin disorder isselected from the group consisting of: psoriasis; atopic dermatitis;allergic contact dermatitis; and alopecia areata.
 3. The method of claim1 wherein the composition is administered by means of intradermalinjection.
 4. The method of claim 1 wherein the composition additionallycomprises an adjuvant.
 5. A method for the treatment of a skin disorder,comprising administering an isolated polypeptide, the polypeptidecomprising an immunogenic portion of an antigen, or a variant thereof,wherein the antigen includes a sequence selected from the groupconsisting of SEQ ID NO: 1-4,9-16, 18-21, 23, 25, 26, 28, 29, 44, 45,47, 52-55, 63, 64, 70, 75, 89, 94, 98, 100-105, 109, 110, 112, 121, 124,125, 134, 135, 140, 141, 143, 145, 147, 152, 154, 156, 158, 160, 165,166, 170, 172, 174, 177, 178, 181, 182, 184, 186, 187, 192 and
 194. 6.The method of claim 5 wherein the skin disorder is selected from thegroup consisting of: psoriasis; atopic dermatitis; allergic contactdermatitis; and alopecia areata.
 7. A method for the treatment of a skindisorder, comprising administering a DNA molecule encoding an isolatedpolypeptide, the polypeptide comprising an immunogenic portion of anantigen, or a variant thereof, wherein the antigen includes a sequenceselected from the group consisting of SEQ ID NO: 1-4,9-16, 18-21, 23,25, 26, 28, 29, 44, 45, 47, 52-55, 63, 64, 70, 75, 89, 94, 98, 100-105,109, 110, 112, 121, 124, 125, 134, 135, 140, 141, 143, 145, 147, 152,154, 156, 158, 160, 165, 166, 170, 172, 174, 177, 178, 181, 182, 184,186, 187, 192 and
 194. 8. The method of claim 7 wherein the skindisorder is selected from the group consisting of: psoriasis; atopicdermatitis; allergic contact dermatitis; and alopecia areata.
 9. Amethod for the treatment of a skin disorder, comprising administering afirst dose of a composition at a first point in time and administering asecond dose of the composition at a second, subsequent point in timewherein the composition comprises a constituent present in or derivedfrom a component selected from the group consisting of: (a) M. vaccaecells; and (b) M. vaccae culture filtrate, the constituent havingantigenic or adjuvant properties.
 10. The method of claim 9 wherein theskin disorder is selected from the group consisting of: psoriasis;atopic dermatitis; allergic contact dermatitis; and alopecia areata. 11.A method for the treatment of a skin disorder, comprising administeringa fusion protein comprising at least one isolated polypeptide includingan immunogenic portion of an antigen, or a variant thereof, wherein theantigen comprises a sequence selected from the group consisting of SEQID NO: 1-4,9-16, 18-21, 23, 25, 26, 28, 29, 44, 45, 47, 52-55, 63, 64,70, 75, 89, 94, 98, 100-105, 109, 110, 112, 121, 124, 125, 134, 135,140, 141, 143, 145, 147, 152, 154, 156, 158, 160, 165, 166, 170, 172,174, 177, 178, 181, 182, 184, 186, 187, 192 and
 194. 12. The method ofclaim 11, wherein the skin disorder is selected from the groupconsisting of: psoriasis; atopic dermatitis; allergic contactdermatitis; and alopecia areata.
 13. A method for inhibiting a Th2immune response comprising administering a composition comprisingdelipidated and deglycolipidated M. vaccae cells.
 14. A method forstimulating the production of IL-10 comprising administering acomposition comprising delipidated and deglycolipidated M. vaccae cells.15. A method for the treatment of psoriatic arthritis, comprisingadministering a composition comprising at least one component selectedfrom the group consisting of (a) inactivated M. vaccae cells; (b) M.vaccae culture filtrate; and (c) delipidated and deglycolipidated M.vaccae cells.
 16. The method of claim 15, wherein the composition isadministered by means of intradermal injection.
 17. The method of claim15, wherein the composition additionally comprises an adjuvant.